Use of TNFα inhibitor for treatment of psoriasis

ABSTRACT

The invention describes methods of treating erosive polyarthritis comprising administering a TNFα antibody, or antigen-binding portion thereof. The invention also describes a method for testing the efficacy of a TNFα antibody, or antigen-binding portion thereof, for the treatment of erosive polyarthritis.

RELATED APPLICATIONS

This application claims priority to U.S. patent application Ser. No.14/266,598, filed on Apr. 30, 2014, which claim priority to U.S. patentapplication Ser. No. 14/228,791, filed on Mar. 28, 2014, which claimspriority to U.S. patent application Ser. No. 11/435,844, filed on May16, 2006, which claims priority to U.S. Provisional Appln. No.60/681,645, which was filed on May 16, 2005. This application alsoclaims priority to U.S. patent application Ser. No. 11/104,117, filed onApr. 11, 2005, which claims priority to U.S. Provisional Appln. No.60/569,100, filed on May 7, 2004, U.S. Provisional Appln. No.60/561,710, filed on Apr. 12, 2004, and U.S. Provisional Appln. No.60/561,139, filed on Apr. 9, 2004.

This application is related to U.S. Pat. Nos. 6,090,382, 6,258,562, and6,509,015. This application is also related to U.S. patent applicationSer. No. 09/801,185, filed Mar. 7, 2001; U.S. patent application Ser.No. 10/163,657, filed Jun. 5, 2002; and U.S. patent application Ser. No.10/422,287, filed Apr. 26, 2002; U.S. patent application Ser. No.10/525,292, filed Aug. 16, 2002; U.S. patent application Ser. No.10/693,233, filed Oct. 24, 2003; U.S. patent application Ser. No.10/622,932, filed Jul. 18, 2003; U.S. patent application Ser. No.10/623,039, filed Jul. 18, 2003; U.S. patent application Ser. No.10/623,076, filed Jul. 18, 2003; U.S. patent application Ser. No.10/623,065, filed Jul. 18, 2003; U.S. patent application Ser. No.10/622,928, filed Jul. 18, 2003; U.S. patent application Ser. No.10/623,075, filed Jul. 18, 2003; U.S. patent application Ser. No.10/623,035, filed Jul. 18, 2003; U.S. patent application Ser. No.10/622,683, filed Jul. 18, 2003; U.S. patent application Ser. No.10/622,205, filed Jul. 18, 2003; U.S. patent application Ser. No.10/622,210, filed Jul. 18, 2003; and U.S. patent application Ser. No.10/623,318, filed Jul. 18, 2003. This application is also related toPCT/US05/12007, filed Apr. 11, 2005. The entire contents of each ofthese patents and patent applications are hereby incorporated herein byreference. The International Patent Application No. PCT/US05/12007corresponds to U.S. Patent Application Publication No. 2006/0009385, andthe entire content of U.S. Patent Application Publication No.2006/0009385 (excluding the claims) is incorporated herein by reference.

BACKGROUND OF THE INVENTION

Polyarthritis may be erosive or non-erosive. In the erosive form, theunderlying disease process erodes the cartilage; in the non-erosiveform, the cartilage is not affected. Erosive polyarthritis is aninflammatory disease of joints that results in tissue destruction anderosion within the affected joint. Erosive polyarthritis occurs in manypatients having inflammatory disorders, including psoriatic arthritis,spondylarthropathies, such as ankylosing spondylitis, and juvenilerheumatoid arthritis. Many of the current treatments of disorders inwhich erosive polyarthritis is a manifestation fail to focus ondecreasing radiographic progression of joint disease.

SUMMARY OF THE INVENTION

There is a need to treat erosive polyarthritis in a safe and effectivemanner. While traditional treatments of erosive polyarthritis, such asadministration of DMARDs, may delay disease progression, traditionaltreatments may be slow to become effective, may lose efficacy with time,and may be associated with potentially serious toxic effects. Thepresent invention provides a safe and effective means for treatingerosive polyarthritis and slowing the progression of joint disease.

The present invention includes methods of treating erosive polyarthritiscomprising administering TNF inhibitors. The invention also provides amethod for treating a human subject suffering from erosivepolyarthritis, comprising administering to the subject an anti-TNFαantibody, such that erosive polyarthritis is treated. Kits and articlesof manufacture comprising a TNFα inhibitor are also included in theinvention.

In one embodiment, the TNFα inhibitor is selected from the groupconsisting of an anti-TNFα antibody, or an antigen-binding portionthereof, a TNF fusion protein, or a recombinant TNF binding protein. Inone embodiment, the TNF fusion protein is etanercept. In anotherembodiment, the anti-TNFα antibody, or antigen-binding portion thereof,is an antibody selected from the group consisting of a humanizedantibody, a chimeric antibody, and a multivalent antibody. In oneembodiment, the anti-TNFα antibody, or antigen-binding portion thereof,is infliximab, golimumab, or adalimumab. In still another embodiment,the anti-TNFα antibody, or antigen-binding portion thereof, is a humanantibody.

The invention provides a method for treating a human subject sufferingfrom erosive polyarthritis, comprising administering to the subject aTNFα antibody, or antigen-binding portion thereof, such that erosivepolyarthritis is treated.

In one embodiment, the TNFα antibody, or antigen-binding portionthereof, is an antibody selected from the group consisting of ahumanized antibody, a chimeric antibody, and a multivalent antibody. Inanother embodiment, the TNFα antibody, or antigen-binding portionthereof, is infliximab or golimumab.

In one embodiment, the TNFα antibody, or antigen-binding portionthereof, is a human antibody. In one embodiment, the human antibody, oran antigen-binding portion thereof, dissociates from human TNFα with aK_(d) of 1×10⁻⁸ M or less and a K_(off) rate constant of 1×10⁻³ s⁻¹ orless, both determined by surface plasmon resonance, and neutralizeshuman TNFα cytotoxicity in a standard in vitro L929 assay with an IC₅₀of 1×10⁻⁷ M or less. In another embodiment, the human antibody, or anantigen-binding portion thereof, has the following characteristics:

a) dissociates from human TNFα with a K_(off) rate constant of 1×10⁻³s⁻¹ or less, as determined by surface plasmon resonance;

b) has a light chain CDR3 domain comprising the amino acid sequence ofSEQ ID NO: 3, or modified from SEQ ID NO: 3 by a single alaninesubstitution at position 1, 4, 5, 7 or 8 or by one to five conservativeamino acid substitutions at positions 1, 3, 4, 6, 7, 8 and/or 9;

c) has a heavy chain CDR3 domain comprising the amino acid sequence ofSEQ ID NO: 4, or modified from SEQ ID NO: 4 by a single alaninesubstitution at position 2, 3, 4, 5, 6, 8, 9, 10 or 11 or by one to fiveconservative amino acid substitutions at positions 2, 3, 4, 5, 6, 8, 9,10, 11 and/or 12. In still another embodiment, the human antibody, or anantigen-binding portion thereof, comprises a light chain variable region(LCVR) having a CDR3 domain comprising the amino acid sequence of SEQ IDNO: 3, or modified from SEQ ID NO: 3 by a single alanine substitution atposition 1, 4, 5, 7 or 8, and comprises a heavy chain variable region(HCVR) having a CDR3 domain comprising the amino acid sequence of SEQ IDNO: 4, or modified from SEQ ID NO: 4 by a single alanine substitution atposition 2, 3, 4, 5, 6, 8, 9, 10 or 11. In yet another embodiment, thehuman antibody, or an antigen-binding portion thereof, comprises a lightchain variable region (LCVR) comprising the amino acid sequence of SEQID NO: 1 and a heavy chain variable region (HCVR) comprising the aminoacid sequence of SEQ ID NO: 2. In one embodiment, the human antibody, oran antigen-binding portion thereof, is adalimumab.

In one embodiment, the TNFα antibody, or antigen-binding portionthereof, is administered to the subject on a biweekly dosing regimen.

In one embodiment, the subject has a disorder in which TNFα activity isdetrimental. In one embodiment, the disorder in which TNFα activity isdetrimental is selected from the group consisting of psoriaticarthritis, ankylosing spondylitis, and juvenile rheumatoid arthritis. Inanother embodiment, the disorder in which TNFα activity is detrimentalis psoriatic arthritis. In still another embodiment, the disorder inwhich TNFα activity is detrimental is rheumatoid arthritis.

In one embodiment, the invention includes further comprisingadministering an additional therapeutic agent to the subject. In oneembodiment, the additional therapeutic agent is methotrexate. In anotherembodiment, the additional therapeutic agent is a Disease ModifyingAnti-Rheumatic Drug (DMARD) or a Nonsteroidal Antiinflammatory Drug(NSAID) or a steroid, or any combination thereof.

The invention includes a method for testing the efficacy of a TNFαantibody, or antigen-binding portion thereof, for decreasingradiographic progression of joint disease associated with erosivepolyarthritis. In one embodiment, the method for testing the efficacy ofa TNFα antibody, or antigen-binding portion thereof, comprisesdetermining the efficacy of the TNFα antibody, or antigen-bindingportion thereof, using a modified Total Sharp Score (mTSS) of a patientpopulation having joint disease associated with erosive polyarthritisand a mTSS of the patient population following administration of theTNFα antibody, or antigen-binding portion thereof, wherein no change ora decrease in the mTSS indicates that the TNFα antibody, orantigen-binding portion thereof, is efficacious for decreasingradiographic progression of joint disease associated with erosivepolyarthritis. In one embodiment, the decrease in the mTSS is about−0.2.

In one embodiment, the patient population also has a disorder in whichTNFα is detrimental. In one embodiment, the disorder in which TNFαactivity is detrimental is selected from the group consisting ofpsoriatic arthritis, ankylosing spondylitis, and juvenile rheumatoidarthritis.

In one embodiment, the TNFα antibody, or antigen-binding portionthereof, is an antibody selected from the group consisting of ahumanized antibody, a chimeric antibody, and a multivalent antibody. Inone embodiment, the TNFα antibody, or antigen-binding portion thereof,is infliximab or golimumab. In another embodiment, the TNFα antibody, orantigen-binding portion thereof, is a human antibody. In one embodiment,the human antibody, or an antigen-binding portion thereof, dissociatesfrom human TNFα with a K_(d) of 1×10⁻⁸ M or less and a K_(off) rateconstant of 1×10⁻³ s⁻¹ or less, both determined by surface plasmonresonance, and neutralizes human TNFα cytotoxicity in a standard invitro L929 assay with an IC₅₀ of 1×10⁻⁷ M or less.

In another embodiment, the human antibody, or an antigen-binding portionthereof, has the following characteristics:

a) dissociates from human TNFα with a K_(off) rate constant of 1×10⁻³s⁻¹ or less, as determined by surface plasmon resonance;

b) has a light chain CDR3 domain comprising the amino acid sequence ofSEQ ID NO: 3, or modified from SEQ ID NO: 3 by a single alaninesubstitution at position 1, 4, 5, 7 or 8 or by one to five conservativeamino acid substitutions at positions 1, 3, 4, 6, 7, 8 and/or 9;

c) has a heavy chain CDR3 domain comprising the amino acid sequence ofSEQ ID NO: 4, or modified from SEQ ID NO: 4 by a single alaninesubstitution at position 2, 3, 4, 5, 6, 8, 9, 10 or 11 or by one to fiveconservative amino acid substitutions at positions 2, 3, 4, 5, 6, 8, 9,10, 11 and/or 12. In still another embodiment, the human antibody, or anantigen-binding portion thereof, comprises a light chain variable region(LCVR) having a CDR3 domain comprising the amino acid sequence of SEQ IDNO: 3, or modified from SEQ ID NO: 3 by a single alanine substitution atposition 1, 4, 5, 7 or 8, and comprises a heavy chain variable region(HCVR) having a CDR3 domain comprising the amino acid sequence of SEQ IDNO: 4, or modified from SEQ ID NO: 4 by a single alanine substitution atposition 2, 3, 4, 5, 6, 8, 9, 10 or 11.

In another embodiment, the human antibody, or an antigen-binding portionthereof, comprises a light chain variable region (LCVR) comprising theamino acid sequence of SEQ ID NO: 1 and a heavy chain variable region(HCVR) comprising the amino acid sequence of SEQ ID NO: 2. In stillanother embodiment of the invention, the human antibody, or anantigen-binding portion thereof, is adalimumab.

In one embodiment of the invention, the TNFα antibody, orantigen-binding portion thereof, is administered to the subject on abiweekly dosing regimen. In one embodiment, antibody, or antigen-bindingportion thereof, is administered in combination with an additionaltherapeutic agent, including, for example methotrexate

The invention describes a method for monitoring the effectiveness of aTNFα antibody, or antigen-binding portion thereof, for the treatment oferosive polyarthritis in a human subject comprising determining theeffectiveness of the TNFα antibody, or antigen-binding portion thereof,using a baseline modified Total Sharp Score (mTSS) of a patientpopulation having erosive polyarthritis and a mTSS score of a patientpopulation following administration of the TNFα antibody, orantigen-binding portion thereof, wherein a result selected from thegroup consisting of a decrease in the mTSS in about 9-27% of the patientpopulation; no change in the mTSS in about 65-73% of the patientpopulation; and an increase in the mTSS in about 9-28% of the patientpopulation, indicates that the TNFα antibody, or antigen-binding portionthereof, is effective at treating erosive polyarthritis.

In one embodiment of the invention, the TNFα antibody, orantigen-binding portion thereof, is an antibody selected from the groupconsisting of a humanized antibody, a chimeric antibody, and amultivalent antibody. In one embodiment, the TNFα antibody, orantigen-binding portion thereof, is infliximab or golimumab. In anotherembodiment, the TNFα antibody, or antigen-binding portion thereof, is ahuman antibody.

In another embodiment, the human antibody, or an antigen-binding portionthereof, dissociates from human TNFα with a K_(d) of 1×10⁻⁸ M or lessand a K_(off) rate constant of 1×10⁻³ s⁻¹ or less, both determined bysurface plasmon resonance, and neutralizes human TNFα cytotoxicity in astandard in vitro L929 assay with an IC₅₀ of 1×10⁻⁷ M or less. In stillanother embodiment, the human antibody, or an antigen-binding portionthereof, is adalimumab.

The invention also includes a method for testing the efficacy of a TNFαantibody, or antigen-binding portion thereof, to treat erosivepolyarthritis associated with psoriatic arthritis, comprisingdetermining the efficacy of the TNFα antibody, or antigen-bindingportion thereof, using a baseline modified Total Sharp Score (mTSS) andeither a baseline Psoriasis Area and Severity Index (PAST) score or abaseline ACR score of a patient population having erosive polyarthritisin comparison with the mTSS and either the PASI or the ACR score of thepatient population following administration of the NFα antibody, orantigen-binding portion thereof, wherein no change or a decrease in themTSS and either an ACR20 response achieved in at least about 57% or aPASI 50 response achieved in at least about 75% of the patientpopulation, indicates that the TNFα antibody, or antigen-binding portionthereof, is efficacious for the treatment of erosive polyarthritisassociated with psoriatic arthritis. In one embodiment, an ACR50response is achieved in at least about 39% of the patient population. Inanother embodiment, an ACR70 response is achieved in at least about 23%of the patient population. In still another embodiment, a PASI75response is achieved in at least about 59% of the patient population. Inyet another embodiment, a PASI90 response is achieved in at least about42% of the patient population. In one embodiment, the TNFα antibody, orantigen-binding portion thereof, is adalimumab.

The invention describes a method for treating erosive polyarthritiscomprising administering to a subject having erosive polyarthritis,adalimumab on a biweekly dosing regimen. In one embodiment, the dose ofadalimumab is about 40 mg.

The invention also includes a kit comprising a pharmaceuticalcomposition comprising a TNFα antibody, or an antigen-binding portionthereof, and a pharmaceutically acceptable carrier, and instructions foradministration of the pharmaceutical composition for the treatment oferosive polyarthritis. In one embodiment, the pharmaceutical compositioncomprises the TNFα antibody, or antigen-binding portion thereof,adalimumab. In one embodiment, pharmaceutical composition comprisesabout 40 mg of adalimumab. In another embodiment, the kit furthercomprises an additional therapeutic agent. In one embodiment, theadditional therapeutic agent is methotrexate.

The invention describes an article of manufacture comprising a packagingmaterial; a TNFα antibody, or antigen-binding portion thereof; and alabel or package insert contained within the packaging materialindicating that the TNFα antibody, or antigen-binding portion thereof,can be used for the treatment of erosive polyarthritis.

The invention also includes an article of manufacture comprising apackaging material; a TNFα antibody, or antigen-binding portion thereof;and a label or package insert contained within the packaging materialindicating that the TNFα antibody, or antigen-binding portion thereof,can be used for to inhibit radiographic progression of joint disease.

In one embodiment, the article of manufacture comprises an antibodyselected from the group consisting of a humanized antibody, a chimericantibody, and a multivalent antibody.

In one embodiment, the TNFα antibody, or antigen-binding portionthereof, is infliximab or golimumab. In another embodiment, the TNFαantibody, or antigen-binding portion thereof, is a human antibody. Inone embodiment, the human antibody, or an antigen-binding portionthereof, dissociates from human TNFα with a K_(d) of 1×10⁻⁸ M or lessand a K_(off) rate constant of 1×10⁻³ s⁻¹ or less, both determined bysurface plasmon resonance, and neutralizes human TNFα cytotoxicity in astandard in vitro L929 assay with an IC₅₀ of 1×10⁻⁷ M or less. Inanother embodiment, the human antibody, or an antigen-binding portionthereof, is adalimumab.

FIGURES

FIGS. 1 a and 1 b show a diagram of the modified total sharp score(mTSS) (FIG. 1 a) and radiographic findings associated with PsA (FIG. 1b).

FIG. 2 shows a cumulative distribution function plot of modified TotalSharps Score (mTSS). The graph shows the change in baseline to Week 24for subjects with both baseline and Week 24 radiographic films.

FIGS. 3 a and 3 b show cumulative distribution function plots of mTSS ofsubjects with (FIG. 3 a) and without (FIG. 3 b) methotrexate (mtx).

FIG. 4 shows a graph of the mean change in mTSS at Week 48.

FIG. 5 shows the percentage of psoriasis patients with ≧PASI 50/75/90response at week 12 following treatment with each multiple variable D2E7dose and the placebo.

FIG. 6 provides results of the mean percentage PASI (Psoriasis Area andSeverity Index) improvement over a 12 week treatment (eow=every otherweek; *=p<0.001 vs. placebo).

FIG. 7 shows a comparative graph of the efficacy response at week 12 andweek 24 for patients with psoriasis and PsA.

FIG. 8 shows a comparative graph of the efficacy response at week 12 andweek 24 for patients with psoriasis and without PsA.

DETAILED DESCRIPTION OF THE INVENTION I. Definitions

In order that the present invention may be more readily understood,certain terms are first defined.

The term “human TNFα” (abbreviated herein as hTNFα, or simply hTNF), asused herein, is intended to refer to a human cytokine that exists as a17 kD secreted form and a 26 kD membrane associated form, thebiologically active form of which is composed of a trimer ofnoncovalently bound 17 kD molecules. The structure of hTNFα is describedfurther in, for example, Pennica, D., et al. (1984) Nature 312:724-729;Davis, J. M., et al. (1987) Biochemistry 26:1322-1326; and Jones, E. Y.,et al. (1989) Nature 338:225-228. The term human TNFα is intended toinclude recombinant human TNFα (rhTNFα), which can be prepared bystandard recombinant expression methods or purchased commercially (R & DSystems, Catalog No. 210-TA, Minneapolis, Minn.). TNFα is also referredto as TNF.

The term “TNFα inhibitor” refers to an agent which interferes with TNFαactivity. The term also includes each of the anti-TNFα human antibodiesand antibody portions described herein as well as those described inU.S. Pat. Nos. 6,090,382; 6,258,562; 6,509,015, and in U.S. patentapplication Ser. Nos. 09/801,185 and 10/302,356. In one embodiment, theTNFα inhibitor used in the invention is an anti-TNFα antibody, or afragment thereof, including infliximab (Remicade®, Johnson and Johnson;described in U.S. Pat. No. 5,656,272, incorporated by reference herein),CDP571 (a humanized monoclonal anti-TNF-alpha IgG4 antibody), CDP 870 (ahumanized monoclonal anti-TNF-alpha antibody fragment), an anti-TNF dAb(Peptech), CNTO 148 (golimumab; Medarex and Centocor, see WO 02/12502),and adalimumab (Humira® Abbott Laboratories, a human anti-TNF mAb,described in U.S. Pat. No. 6,090,382 as D2E7). Additional TNF antibodieswhich can be used in the invention are described in U.S. Pat. Nos.6,593,458; 6,498,237; 6,451,983; and 6,448,380, each of which isincorporated by reference herein. In another embodiment, the TNFαinhibitor is a TNF fusion protein, e.g., etanercept (Enbrel®, Amgen;described in WO 91/03553 and WO 09/406,476, incorporated by referenceherein). In another embodiment, the TNFα inhibitor is a recombinant TNFbinding protein (r-TBP-I) (Serono).

The term “antibody”, as used herein, is intended to refer toimmunoglobulin molecules comprised of four polypeptide chains, two heavy(H) chains and two light (L) chains inter-connected by disulfide bonds.Each heavy chain is comprised of a heavy chain variable region(abbreviated herein as HCVR or VH) and a heavy chain constant region.The heavy chain constant region is comprised of three domains, CH1, CH2and CH3. Each light chain is comprised of a light chain variable region(abbreviated herein as LCVR or VL) and a light chain constant region.The light chain constant region is comprised of one domain, CL. The VHand VL regions can be further subdivided into regions ofhypervariability, termed complementarity determining regions (CDR),interspersed with regions that are more conserved, termed frameworkregions (FR). Each VH and VL is composed of three CDRs and four FRs,arranged from amino-terminus to carboxy-terminus in the following order:FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The antibodies of the inventionare described in further detail in U.S. Pat. Nos. 6,090,382; 6,258,562;and 6,509,015, each of which is incorporated herein by reference in itsentirety.

The term “antigen-binding portion” of an antibody (or simply “antibodyportion”), as used herein, refers to one or more fragments of anantibody that retain the ability to specifically bind to an antigen(e.g., hTNFα). It has been shown that the antigen-binding function of anantibody can be performed by fragments of a full-length antibody.Examples of binding fragments encompassed within the term“antigen-binding portion” of an antibody include (i) a Fab fragment, amonovalent fragment consisting of the VL, VH, CL and CH1 domains; (ii) aF(ab′)₂ fragment, a bivalent fragment comprising two Fab fragmentslinked by a disulfide bridge at the hinge region; (iii) a Fd fragmentconsisting of the VH and CH1 domains; (iv) a Fv fragment consisting ofthe VL and VH domains of a single arm of an antibody, (v) a dAb fragment(Ward et al. (1989) Nature 341:544-546), which consists of a VH domain;and (vi) an isolated complementarity determining region (CDR).Furthermore, although the two domains of the Fv fragment, VL and VH, arecoded for by separate genes, they can be joined, using recombinantmethods, by a synthetic linker that enables them to be made as a singleprotein chain in which the VL and VH regions pair to form monovalentmolecules (known as single chain Fv (scFv); see e.g., Bird et al. (1988)Science 242:423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. USA85:5879-5883). Such single chain antibodies are also intended to beencompassed within the term “antigen-binding portion” of an antibody.Other forms of single chain antibodies, such as diabodies are alsoencompassed. Diabodies are bivalent, bispecific antibodies in which VHand VL domains are expressed on a single polypeptide chain, but using alinker that is too short to allow for pairing between the two domains onthe same chain, thereby forcing the domains to pair with complementarydomains of another chain and creating two antigen binding sites (seee.g., Holliger et al. (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448;Poljak et al. (1994) Structure 2:1121-1123). The antibody portions ofthe invention are described in further detail in U.S. Pat. Nos.6,090,382, 6,258,562, 6,509,015, each of which is incorporated herein byreference in its entirety.

Binding fragments are produced by recombinant DNA techniques, or byenzymatic or chemical cleavage of intact immunoglobulins. Bindingfragments include Fab, Fab′, F(ab′)₂, Fabc, Fv, single chains, andsingle-chain antibodies. Other than “bispecific” or “bifunctional”immunoglobulins or antibodies, an immunoglobulin or antibody isunderstood to have each of its binding sites identical. A “bispecific”or “bifunctional antibody” is an artificial hybrid antibody having twodifferent heavy/light chain pairs and two different binding sites.Bispecific antibodies can be produced by a variety of methods includingfusion of hybridomas or linking of Fab′ fragments. See, e.g.,Songsivilai & Lachmann, Clin. Exp. Immunol. 79:315-321 (1990); Kostelnyet al., J. Immunol. 148, 1547-1553 (1992).

A “conservative amino acid substitution”, as used herein, is one inwhich one amino acid residue is replaced with another amino acid residuehaving a similar side chain. Families of amino acid residues havingsimilar side chains have been defined in the art, including basic sidechains (e.g., lysine, arginine, histidine), acidic side chains (e.g.,aspartic acid, glutamic acid), uncharged polar side chains (e.g.,glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine),nonpolar side chains (e.g., alanine, valine, leucine, isoleucine,proline, phenylalanine, methionine, tryptophan), beta-branched sidechains (e.g., threonine, valine, isoleucine) and aromatic side chains(e.g., tyrosine, phenylalanine, tryptophan, histidine).

The term “human antibody”, as used herein, is intended to includeantibodies having variable and constant regions derived from humangermline immunoglobulin sequences. The human antibodies of the inventionmay include amino acid residues not encoded by human germlineimmunoglobulin sequences (e.g., mutations introduced by random orsite-specific mutagenesis in vitro or by somatic mutation in vivo), forexample in the CDRs and in particular CDR3. However, the term “humanantibody”, as used herein, is not intended to include antibodies inwhich CDR sequences derived from the germline of another mammalianspecies, such as a mouse, have been grafted onto human frameworksequences.

The term “recombinant human antibody”, as used herein, is intended toinclude all human antibodies that are prepared, expressed, created orisolated by recombinant means, such as antibodies expressed using arecombinant expression vector transfected into a host cell (describedfurther below), antibodies isolated from a recombinant, combinatorialhuman antibody library (described further below), antibodies isolatedfrom an animal (e.g., a mouse) that is transgenic for humanimmunoglobulin genes (see e.g., Taylor et al. (1992) Nucl. Acids Res.20:6287) or antibodies prepared, expressed, created or isolated by anyother means that involves splicing of human immunoglobulin genesequences to other DNA sequences. Such recombinant human antibodies havevariable and constant regions derived from human germline immunoglobulinsequences. In certain embodiments, however, such recombinant humanantibodies are subjected to in vitro mutagenesis (or, when an animaltransgenic for human Ig sequences is used, in vivo somatic mutagenesis)and thus the amino acid sequences of the VH and VL regions of therecombinant antibodies are sequences that, while derived from andrelated to human germline VH and VL sequences, may not naturally existwithin the human antibody germline repertoire in vivo.

An “isolated antibody”, as used herein, is intended to refer to anantibody that is substantially free of other antibodies having differentantigenic specificities (e.g., an isolated antibody that specificallybinds hTNFα is substantially free of antibodies that specifically_bindantigens other than hTNFα). An isolated antibody that specifically bindshTNFα may, however, have cross-reactivity to other antigens, such asTNFα molecules from other species (discussed in further detail below).Moreover, an isolated antibody may be substantially free of othercellular material and/or chemicals.

A “neutralizing antibody”, as used herein (or an “antibody thatneutralized hTNFα activity”), is intended to refer to an antibody whosebinding to hTNFα results in inhibition of the biological activity ofhTNFα. This inhibition of the biological activity of hTNFα can beassessed by measuring one or more indicators of hTNFα biologicalactivity, such as hTNFα-induced cytotoxicity (either in vitro or invivo), hTNFα-induced cellular activation and hTNFα binding to hTNFαreceptors. These indicators of hTNFα biological activity can be assessedby one or more of several standard in vitro or in vivo assays known inthe art (see U.S. Pat. No. 6,090,382). Preferably, the ability of anantibody to neutralize hTNFα activity is assessed by inhibition ofhTNFα-induced cytotoxicity of L929 cells. As an additional oralternative parameter of hTNFα activity, the ability of an antibody toinhibit hTNFα-induced expression of ELAM-1 on HUVEC, as a measure ofhTNFα-induced cellular activation, can be assessed.

The term “surface plasmon resonance”, as used herein, refers to anoptical phenomenon that allows for the analysis of real-time biospecificinteractions by detection of alterations in protein concentrationswithin a biosensor matrix, for example using the BIAcore system(Pharmacia Biosensor AB, Uppsala, Sweden and Piscataway, N.J.). Forfurther descriptions, see Example 1 of U.S. Pat. No. 6,258,562 andJönsson et al. (1993) Ann. Biol. Clin. 51:19; Jönsson et al., (1991)Biotechniques 11:620-627; Johnsson et al. (1995) J. Mol. Recognit.8:125; and Johnnson et al. (1991) Anal. Biochem. 198:268.

The term “K_(off)”, as used herein, is intended to refer to the off rateconstant for dissociation of an antibody from the antibody/antigencomplex.

The term “K_(d)”, as used herein, is intended to refer to thedissociation constant of a particular antibody-antigen interaction.

The term “IC₅₀” as used herein, is intended to refer to theconcentration of the inhibitor required to inhibit the biologicalendpoint of interest, e.g., neutralize cytotoxicity activity.

The term “nucleic acid molecule”, as used herein, is intended to includeDNA molecules and RNA molecules. A nucleic acid molecule may besingle-stranded or double-stranded, but preferably is double-strandedDNA.

The term “isolated nucleic acid molecule”, as used herein in referenceto nucleic acids encoding antibodies or antibody portions (e.g., VH, VL,CDR3) that bind hTNFα, is intended to refer to a nucleic acid moleculein which the nucleotide sequences encoding the antibody or antibodyportion are free of other nucleotide sequences encoding antibodies orantibody portions that bind antigens other than hTNFα, which othersequences may naturally flank the nucleic acid in human genomic DNA.Thus, for example, an isolated nucleic acid of the invention encoding aVH region of an anti-hTNFα antibody contains no other sequences encodingother VH regions that bind antigens other than hTNFα.

The term “vector”, as used herein, is intended to refer to a nucleicacid molecule capable of transporting another nucleic acid to which ithas been linked. One type of vector is a “plasmid”, which refers to acircular double stranded DNA loop into which additional DNA segments maybe ligated. Another type of vector is a viral vector, wherein additionalDNA segments may be ligated into the viral genome. Certain vectors arecapable of autonomous replication in a host cell into which they areintroduced (e.g., bacterial vectors having a bacterial origin ofreplication and episomal mammalian vectors). Other vectors (e.g.,non-episomal mammalian vectors) can be integrated into the genome of ahost cell upon introduction into the host cell, and thereby arereplicated along with the host genome. Moreover, certain vectors arecapable of directing the expression of genes to which they areoperatively linked. Such vectors are referred to herein as “recombinantexpression vectors” (or simply, “expression vectors”). In general,expression vectors of utility in recombinant DNA techniques are often inthe form of plasmids. In the present specification, “plasmid” and“vector” may be used interchangeably as the plasmid is the most commonlyused form of vector. However, the invention is intended to include suchother forms of expression vectors, such as viral vectors (e.g.,replication defective retroviruses, adenoviruses and adeno-associatedviruses), which serve equivalent functions.

The term “recombinant host cell” (or simply “host cell”), as usedherein, is intended to refer to a cell into which a recombinantexpression vector has been introduced. It should be understood that suchterms are intended to refer not only to the particular subject cell butto the progeny of such a cell. Because certain modifications may occurin succeeding generations due to either mutation or environmentalinfluences, such progeny may not, in fact, be identical to the parentcell, but are still included within the scope of the term “host cell” asused herein.

The term “dose,” as used herein, refers to an amount of TNFα inhibitorwhich is administered to a subject.

The term “multiple-variable dose” includes different doses of a TNFαinhibitor which are administered to a subject for therapeutic treatment.“Multiple-variable dose regimen” or “multiple-variable dose therapy”describe a treatment schedule which is based on administering differentamounts of TNFα inhibitor at various time points throughout the courseof treatment. In one embodiment, the invention describes amultiple-variable dose method of treatment of erosive polyarthritiscomprising an induction phase and a treatment phase, wherein a TNFαinhibitor is administered at a higher dose during the induction phasethan the treatment phase. Multiple-variable dose regimens are describedin PCT application no. PCT/US05/12007.

The term “dosing”, as used herein, refers to the administration of asubstance (e.g., an anti-TNFα antibody) to achieve a therapeuticobjective (e.g., the treatment of erosive polyarthritis).

The terms “biweekly dosing regimen”, “biweekly dosing”, and “biweeklyadministration”, as used herein, refer to the time course ofadministering a substance (e.g., an anti-TNFα antibody) to a subject toachieve a therapeutic objective (e.g., the treatment of erosivepolyarthritis). The biweekly dosing regimen is not intended to include aweekly dosing regimen. Preferably, the substance is administered every9-19 days, more preferably, every 11-17 days, even more preferably,every 13-15 days, and most preferably, every 14 days.

The term “combination” as in the phrase “a first agent in combinationwith a second agent” includes co-administration of a first agent and asecond agent, which for example may be dissolved or intermixed in thesame pharmaceutically acceptable carrier, or administration of a firstagent, followed by the second agent, or administration of the secondagent, followed by the first agent. The present invention, therefore,includes methods of combination therapeutic treatment and combinationpharmaceutical compositions. In one embodiment, the invention provides acombination therapy for treating erosive polyarthritis comprisingadministering an anti-TNF antibody.

The term “concomitant” as in the phrase “concomitant therapeutictreatment” includes administering an agent in the presence of a secondagent. A concomitant therapeutic treatment method includes methods inwhich the first, second, third, or additional agents areco-administered. A concomitant therapeutic treatment method alsoincludes methods in which the first or additional agents areadministered in the presence of a second or additional agents, whereinthe second or additional agents, for example, may have been previouslyadministered. A concomitant therapeutic treatment method may be executedstep-wise by different actors. For example, one actor may administer toa subject a first agent and a second actor may to administer to thesubject a second agent, and the administering steps may be executed atthe same time, or nearly the same time, or at distant times, so long asthe first agent (and additional agents) are after administration in thepresence of the second agent (and additional agents). The actor and thesubject may be the same entity (e.g., human).

The term “combination therapy”, as used herein, refers to theadministration of two or more therapeutic substances, e.g., an anti-TNFαantibody and another drug. The other drug(s) may be administeredconcomitant with, prior to, or following the administration of ananti-TNFα antibody.

The term “kit” as used herein refers to a packaged product or article ofmanufacture comprising components. The kit preferably comprises a box orcontainer that holds the components of the kit. The box or container isaffixed with a label or a Food and Drug Administration approvedprotocol. The box or container holds components of the invention whichare preferably contained within plastic, polyethylene, polypropylene,ethylene, or propylene vessels. The vessels can be capped-tubes orbottles. The kit can also include instructions for administering theTNFα antibody, or antigen-binding portion thereof. In one embodiment thekit of the invention includes the formulation comprising the humanantibody D2E7, as described in PCT/IB03/04502 and U.S. application Ser.No. 10/222,140.

Various aspects of the invention are described in further detail herein.

II. TNFα Inhibitors

This invention provides a method of treating erosive polyarthritis inwhich the administration of a TNFα inhibitor e.g., a TNFα antibody, orantigen-binding portion thereof, is beneficial. In one embodiment, thesemethods include administration of isolated human antibodies, orantigen-binding portions thereof, that bind to human TNFα with highaffinity and a low off rate, and have a high neutralizing capacity.Preferably, the human antibodies of the invention are recombinant,neutralizing human anti-hTNFα antibodies.

In one embodiment, the TNFα inhibitor used in the invention is ananti-TNFα antibody, or a fragment thereof, including infliximab(Remicade®, Johnson and Johnson; described in U.S. Pat. No. 5,656,272,incorporated by reference herein), CDP571 (a humanized monoclonalanti-TNF-alpha IgG4 antibody), CDP 870 (a humanized monoclonalanti-TNF-alpha antibody fragment), an anti-TNF dAb (Peptech), CNTO 148(golimumab; Medarex and Centocor, see WO 02/12502), and adalimumab(Humira® Abbott Laboratories, a human anti-TNF mAb, described in U.S.Pat. No. 6,090,382 as D2E7). Additional TNF antibodies which can be usedin the invention are described in U.S. Pat. Nos. 6,593,458; 6,498,237;6,451,983; and 6,448,380, each of which is incorporated by referenceherein.

The most preferred recombinant, neutralizing antibody used in theinvention is referred to herein as D2E7, also referred to as HUMIRA® andadalimumab (the amino acid sequence of the D2E7 VL region is shown inSEQ ID NO: 1; the amino acid sequence of the D2E7 VH region is shown inSEQ ID NO: 2). The properties of D2E7 (HUMIRA®) have been described inSalfeld et al., U.S. Pat. Nos. 6,090,382, 6,258,562, and 6,509,015,which are each incorporated by reference herein. Other examples of TNFαinhibitors include chimeric and humanized murine anti-hTNFα antibodieswhich have undergone clinical testing for treatment of rheumatoidarthritis (see e.g., Elliott et al. (1994) Lancet 344:1125-1127; Elliotet al. (1994) Lancet 344:1105-1110; Rankin et al., (1995) Br. J.Rheumatol. 34:334-342). In another embodiment, the anti-TNFα antibody ismultivalent.

In one embodiment, the method of treating erosive polyarthritis of theinvention includes the administration of D2E7 antibodies and antibodyportions, D2E7-related antibodies and antibody portions, and other humanantibodies and antibody portions with equivalent properties to D2E7,such as high affinity binding to hTNFα with low dissociation kineticsand high neutralizing capacity. In one embodiment, the inventionprovides a method for treating erosive polyarthritis with an isolatedhuman antibody, or an antigen-binding portion thereof, that dissociatesfrom human TNFα with a K_(d) of 1×10⁻⁸ M or less and a K_(off) rateconstant of 1×10⁻³ s⁻¹ or less, both determined by surface plasmonresonance, and neutralizes human TNFα cytotoxicity in a standard invitro L929 assay with an IC₅₀ of 1×10⁻⁷ M or less. More preferably, theisolated human antibody, or antigen-binding portion thereof, dissociatesfrom human TNFα with a K_(off) of 5×10⁻⁴ s⁻¹ or less, or even morepreferably, with a K_(off) of 1×10⁻⁴ s⁻¹ or less. More preferably, theisolated human antibody, or antigen-binding portion thereof, neutralizeshuman TNFα cytotoxicity in a standard in vitro L929 assay with an IC₅₀of 1×10⁻⁸ M or less, even more preferably with an IC₅₀ of 1×10⁻⁹ M orless and still more preferably with an IC₅₀ of 1×10⁻¹⁰ M or less. In apreferred embodiment, the antibody is an isolated human recombinantantibody, or an antigen-binding portion thereof.

It is well known in the art that antibody heavy and light chain CDR3domains play an important role in the binding specificity/affinity of anantibody for an antigen. Accordingly, in another aspect, the inventionpertains to methods of treating erosive polyarthritis by administeringhuman antibodies that have slow dissociation kinetics for associationwith hTNFα and that have light and heavy chain CDR3 domains thatstructurally are identical to or related to those of D2E7. Position 9 ofthe D2E7 VL CDR3 can be occupied by Ala or Thr without substantiallyaffecting the K_(off). Accordingly, a consensus motif for the D2E7 VLCDR3 comprises the amino acid sequence: Q-R-Y-N-R-A-P-Y-(T/A) (SEQ IDNO: 3). Additionally, position 12 of the D2E7 VH CDR3 can be occupied byTyr or Asn, without substantially affecting the K_(off). Accordingly, aconsensus motif for the D2E7 VH CDR3 comprises the amino acid sequence:V-S-Y-L-S-T-A-S-S-L-D-(Y/N) (SEQ ID NO: 4). Moreover, as demonstrated inExample 2 of U.S. Pat. No. 6,090,382, the CDR3 domain of the D2E7 heavyand light chains is amenable to substitution with a single alanineresidue (at position 1, 4, 5, 7 or 8 within the VL CDR3 or at position2, 3, 4, 5, 6, 8, 9, 10 or 11 within the VH CDR3) without substantiallyaffecting the K_(off) Still further, the skilled artisan will appreciatethat, given the amenability of the D2E7 VL and VH CDR3 domains tosubstitutions by alanine, substitution of other amino acids within theCDR3 domains may be possible while still retaining the low off rateconstant of the antibody, in particular substitutions with conservativeamino acids. Preferably, no more than one to five conservative aminoacid substitutions are made within the D2E7 VL and/or VH CDR3 domains.More preferably, no more than one to three conservative amino acidsubstitutions are made within the D2E7 VL and/or VH CDR3 domains.Additionally, conservative amino acid substitutions should not be madeat amino acid positions critical for binding to hTNFα. Positions 2 and 5of the D2E7 VL CDR3 and positions 1 and 7 of the D2E7 VH CDR3 appear tobe critical for interaction with hTNFα and thus, conservative amino acidsubstitutions preferably are not made at these positions (although analanine substitution at position 5 of the D2E7 VL CDR3 is acceptable, asdescribed above) (see U.S. Pat. No. 6,090,382).

Accordingly, in another embodiment, the invention provides methods oftreating erosive polyarthritis by the administration of an isolatedhuman antibody, or antigen-binding portion thereof. The antibody orantigen-binding portion thereof preferably contains the followingcharacteristics:

a) dissociates from human TNFα with a K_(off) rate constant of 1×10⁻³s⁻¹ or less, as determined by surface plasmon resonance;

b) has a light chain CDR3 domain comprising the amino acid sequence ofSEQ ID NO: 3, or modified from SEQ ID NO: 3 by a single alaninesubstitution at position 1, 4, 5, 7 or 8 or by one to five conservativeamino acid substitutions at positions 1, 3, 4, 6, 7, 8 and/or 9;

c) has a heavy chain CDR3 domain comprising the amino acid sequence ofSEQ ID NO: 4, or modified from SEQ ID NO: 4 by a single alaninesubstitution at position 2, 3, 4, 5, 6, 8, 9, 10 or 11 or by one to fiveconservative amino acid substitutions at positions 2, 3, 4, 5, 6, 8, 9,10, 11 and/or 12.

More preferably, the antibody, or antigen-binding portion thereof,dissociates from human TNFα with a K_(off) of 5×10⁻⁴ s⁻¹ or less. Evenmore preferably, the antibody, or antigen-binding portion thereof,dissociates from human TNFα with a K_(off) of 1×10⁻⁴ s⁻¹ or less.

In yet another embodiment, the invention provides a method of treatingerosive polyarthritis by administration of an isolated human antibody,or antigen-binding portion thereof. The antibody or antigen-bindingportion thereof preferably contains a light chain variable region (LCVR)having a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 3,or modified from SEQ ID NO: 3 by a single alanine substitution atposition 1, 4, 5, 7 or 8, and with a heavy chain variable region (HCVR)having a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 4,or modified from SEQ ID NO: 4 by a single alanine substitution atposition 2, 3, 4, 5, 6, 8, 9, 10 or 11. Preferably, the LCVR further hasa CDR2 domain comprising the amino acid sequence of SEQ ID NO: 5 (i.e.,the D2E7 VL CDR2) and the HCVR further has a CDR2 domain comprising theamino acid sequence of SEQ ID NO: 6 (i.e., the D2E7 VH CDR2). Even morepreferably, the LCVR further has CDR1 domain comprising the amino acidsequence of SEQ ID NO: 7 (i.e., the D2E7 VL CDR1) and the HCVR has aCDR1 domain comprising the amino acid sequence of SEQ ID NO: 8 (i.e.,the D2E7 VH CDR1). The framework regions for VL preferably are from theV_(κ)I human germline family, more preferably from the A20 humangermline Vk gene and most preferably from the D2E7 VL frameworksequences shown in FIGS. 1A and 1B of U.S. Pat. No. 6,090,382. Theframework regions for VH preferably are from the V_(H)3 human germlinefamily, more preferably from the DP-31 human germline VH gene and mostpreferably from the D2E7 VH framework sequences shown in FIGS. 2A and 2Bof U.S. Pat. No. 6,090,382.

Accordingly, in another embodiment, the invention provides a method oftreating erosive polyarthritis by the administration of an isolatedhuman antibody, or antigen-binding portion thereof. The antibody orantigen-binding portion thereof preferably contains a light chainvariable region (LCVR) comprising the amino acid sequence of SEQ ID NO:1 (i.e., the D2E7 VL) and a heavy chain variable region (HCVR)comprising the amino acid sequence of SEQ ID NO: 2 (i.e., the D2E7 VH).In certain embodiments, the antibody comprises a heavy chain constantregion, such as an IgG1, IgG2, IgG3, IgG4, IgA, IgE, IgM or IgD constantregion. Preferably, the heavy chain constant region is an IgG1 heavychain constant region or an IgG4 heavy chain constant region.Furthermore, the antibody can comprise a light chain constant region,either a kappa light chain constant region or a lambda light chainconstant region. Preferably, the antibody comprises a kappa light chainconstant region. Alternatively, the antibody portion can be, forexample, a Fab fragment or a single chain Fv fragment.

In still other embodiments, the invention describes a method of treatingerosive polyarthritis in which the administration of an anti-TNFαantibody wherein the antibody is an isolated human antibody, or anantigen-binding portion thereof. The antibody or antigen-binding portionthereof preferably contains D2E7-related VL and VH CDR3 domains, forexample, antibodies, or antigen-binding portions thereof, with a lightchain variable region (LCVR) having a CDR3 domain comprising an aminoacid sequence selected from the group consisting of SEQ ID NO: 3, SEQ IDNO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20,SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO:25 and SEQ ID NO: 26 or with a heavy chain variable region (HCVR) havinga CDR3 domain comprising an amino acid sequence selected from the groupconsisting of SEQ ID NO: 4, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29,SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO:34 and SEQ ID NO: 35.

The TNFα antibody used in the invention can also be modified. In someembodiments, the TNFα antibody or antigen binding fragments thereof, ischemically modified to provide a desired effect. For example, pegylationof antibodies and antibody fragments of the invention may be carried outby any of the pegylation reactions known in the art, as described, forexample, in the following references: Focus on Growth Factors 3:4-10(1992); EP 0 154 316; and EP 0 401 384 (each of which is incorporated byreference herein in its entirety). Preferably, the pegylation is carriedout via an acylation reaction or an alkylation reaction with a reactivepolyethylene glycol molecule (or an analogous reactive water-solublepolymer). A preferred water-soluble polymer for pegylation of theantibodies and antibody fragments of the invention is polyethyleneglycol (PEG). As used herein, “polyethylene glycol” is meant toencompass any of the forms of PEG that have been used to derivatizeother proteins, such as mono (Cl—ClO) alkoxy- or aryloxy-polyethyleneglycol.

Methods for preparing pegylated antibodies and antibody fragments of theinvention will generally comprise the steps of (a) reacting the antibodyor antibody fragment with polyethylene glycol, such as a reactive esteror aldehyde derivative of PEG, under conditions whereby the antibody orantibody fragment becomes attached to one or more PEG groups, and (b)obtaining the reaction products. It will be apparent to one of ordinaryskill in the art to select the optimal reaction conditions or theacylation reactions based on known parameters and the desired result.

Pegylated antibodies and antibody fragments may generally be used totreat erosive polyarthritis and TNFα-related disorders of the inventionby administration of the TNFα antibodies and antibody fragmentsdescribed herein. Generally the pegylated antibodies and antibodyfragments have increased half-life, as compared to the nonpegylatedantibodies and antibody fragments. The pegylated antibodies and antibodyfragments may be employed alone, together, or in combination with otherpharmaceutical compositions.

In yet another embodiment of the invention, TNFα antibodies or fragmentsthereof can be altered wherein the constant region of the antibody ismodified to reduce at least one constant region-mediated biologicaleffector function relative to an unmodified antibody. To modify anantibody of the invention such that it exhibits reduced binding to theFc receptor, the immunoglobulin constant region segment of the antibodycan be mutated at particular regions necessary for Fc receptor (FcR)interactions (see e.g., Canfield and Morrison (1991) J. Exp. Med.173:1483-1491; and Lund et al. (1991) J. of Immunol. 147:2657-2662).Reduction in FcR binding ability of the antibody may also reduce othereffector functions which rely on FcR interactions, such as opsonizationand phagocytosis and antigen-dependent cellular cytotoxicity.

An antibody or antibody portion of the invention can be derivatized orlinked to another functional molecule (e.g., another peptide orprotein). Accordingly, the antibodies and antibody portions of theinvention are intended to include derivatized and otherwise modifiedforms of the human anti-hTNFα antibodies described herein, includingimmunoadhesion molecules. For example, an antibody or antibody portionof the invention can be functionally linked (by chemical coupling,genetic fusion, noncovalent association or otherwise) to one or moreother molecular entities, such as another antibody (e.g., a bispecificantibody or a diabody), a detectable agent, a cytotoxic agent, apharmaceutical agent, and/or a protein or peptide that can mediateassociate of the antibody or antibody portion with another molecule(such as a streptavidin core region or a polyhistidine tag).

One type of derivatized antibody is produced by crosslinking two or moreantibodies (of the same type or of different types, e.g., to createbispecific antibodies). Suitable crosslinkers include those that areheterobifunctional, having two distinctly reactive groups separated byan appropriate spacer (e.g., m-maleimidobenzoyl-N-hydroxysuccinimideester) or homobifunctional (e.g., disuccinimidyl suberate). Such linkersare available from Pierce Chemical Company, Rockford, Ill.

Useful detectable agents with which an antibody or antibody portion ofthe invention may be derivatized include fluorescent compounds.Exemplary fluorescent detectable agents include fluorescein, fluoresceinisothiocyanate, rhodamine, 5-dimethylamine-1-napthalenesulfonylchloride, phycoerythrin and the like. An antibody may also bederivatized with detectable enzymes, such as alkaline phosphatase,horseradish peroxidase, glucose oxidase and the like. When an antibodyis derivatized with a detectable enzyme, it is detected by addingadditional reagents that the enzyme uses to produce a detectablereaction product. For example, when the detectable agent horseradishperoxidase is present, the addition of hydrogen peroxide anddiaminobenzidine leads to a colored reaction product, which isdetectable. An antibody may also be derivatized with biotin, anddetected through indirect measurement of avidin or streptavidin binding.

An antibody, or antibody portion, of the invention can be prepared byrecombinant expression of immunoglobulin light and heavy chain genes ina host cell. To express an antibody recombinantly, a host cell istransfected with one or more recombinant expression vectors carrying DNAfragments encoding the immunoglobulin light and heavy chains of theantibody such that the light and heavy chains are expressed in the hostcell and, preferably, secreted into the medium in which the host cellsare cultured, from which medium the antibodies can be recovered.Standard recombinant DNA methodologies are used to obtain antibody heavyand light chain genes, incorporate these genes into recombinantexpression vectors and introduce the vectors into host cells, such asthose described in Sambrook, Fritsch and Maniatis (eds), MolecularCloning; A Laboratory Manual, Second Edition, Cold Spring Harbor, N.Y.,(1989), Ausubel et al. (eds.) Current Protocols in Molecular Biology,Greene Publishing Associates, (1989) and in U.S. Pat. No. 4,816,397 byBoss et al.

To express D2E7 or a D2E7-related antibody, DNA fragments encoding thelight and heavy chain variable regions are first obtained. These DNAscan be obtained by amplification and modification of germline light andheavy chain variable sequences using the polymerase chain reaction(PCR). Germline DNA sequences for human heavy and light chain variableregion genes are known in the art (see e.g., the “Vbase” human germlinesequence database; see also Kabat et al. (1991) Sequences of Proteins ofImmunological Interest, Fifth Edition, U.S. Department of Health andHuman Services, NIH Publication No. 91-3242; Tomlinson et al. (1992)“The Repertoire of Human Germline V_(H) Sequences Reveals about FiftyGroups of V_(H) Segments with Different Hypervariable Loops” J. Mol.Biol. 227:776-798; and Cox et al. (1994) “A Directory of Human Germ-lineV₇₈ Segments Reveals a Strong Bias in their Usage” Eur. J. Immunol.24:827-836; the contents of each of which are expressly incorporatedherein by reference). To obtain a DNA fragment encoding the heavy chainvariable region of D2E7, or a D2E7-related antibody, a member of theV_(H)3 family of human germline VH genes is amplified by standard PCR.Most preferably, the DP-31 VH germline sequence is amplified. To obtaina DNA fragment encoding the light chain variable region of D2E7, or aD2E7-related antibody, a member of the V_(κ)I family of human germlineVL genes is amplified by standard PCR. Most preferably, the A20 VLgermline sequence is amplified. PCR primers suitable for use inamplifying the DP-31 germline VH and A20 germline VL sequences can bedesigned based on the nucleotide sequences disclosed in the referencescited supra, using standard methods.

Once the germline VH and VL fragments are obtained, these sequences canbe mutated to encode the D2E7 or D2E7-related amino acid sequencesdisclosed herein. The amino acid sequences encoded by the germline VHand VL DNA sequences are first compared to the D2E7 or D2E7-related VHand VL amino acid sequences to identify amino acid residues in the D2E7or D2E7-related sequence that differ from germline. Then, theappropriate nucleotides of the germline DNA sequences are mutated suchthat the mutated germline sequence encodes the D2E7 or D2E7-relatedamino acid sequence, using the genetic code to determine whichnucleotide changes should be made. Mutagenesis of the germline sequencesis carried out by standard methods, such as PCR-mediated mutagenesis (inwhich the mutated nucleotides are incorporated into the PCR primers suchthat the PCR product contains the mutations) or site-directedmutagenesis.

Once DNA fragments encoding D2E7 or D2E7-related VH and VL segments areobtained (by amplification and mutagenesis of germline VH and VL genes,as described above), these DNA fragments can be further manipulated bystandard recombinant DNA techniques, for example to convert the variableregion genes to full-length antibody chain genes, to Fab fragment genesor to a scFv gene. In these manipulations, a VL- or VH-encoding DNAfragment is operatively linked to another DNA fragment encoding anotherprotein, such as an antibody constant region or a flexible linker. Theterm “operatively linked”, as used in this context, is intended to meanthat the two DNA fragments are joined such that the amino acid sequencesencoded by the two DNA fragments remain in-frame.

The isolated DNA encoding the VH region can be converted to afull-length heavy chain gene by operatively linking the VH-encoding DNAto another DNA molecule encoding heavy chain constant regions (CH1, CH2and CH3). The sequences of human heavy chain constant region genes areknown in the art (see e.g., Kabat et al. (1991) Sequences of Proteins ofImmunological Interest, Fifth Edition, U.S. Department of Health andHuman Services, NIH Publication No. 91-3242) and DNA fragmentsencompassing these regions can be obtained by standard PCRamplification. The heavy chain constant region can be an IgG1, IgG2,IgG3, IgG4, IgA, IgE, IgM or IgD constant region, but most preferably isan IgG1 or IgG4 constant region. For a Fab fragment heavy chain gene,the VH-encoding DNA can be operatively linked to another DNA moleculeencoding only the heavy chain CH₁ constant region.

The isolated DNA encoding the VL region can be converted to afull-length light chain gene (as well as a Fab light chain gene) byoperatively linking the VL-encoding DNA to another DNA molecule encodingthe light chain constant region, CL. The sequences of human light chainconstant region genes are known in the art (see e.g., Kabat et al.(1991) Sequences of Proteins of Immunological Interest, Fifth Edition,U.S. Department of Health and Human Services, NIH Publication No.91-3242) and DNA fragments encompassing these regions can be obtained bystandard PCR amplification. The light chain constant region can be akappa or lambda constant region, but most preferably is a kappa constantregion.

To create a scFv gene, the VH- and VL-encoding DNA fragments areoperatively linked to another fragment encoding a flexible linker, e.g.,encoding the amino acid sequence (Gly₄-Ser)₃, such that the VH and VLsequences can be expressed as a contiguous single-chain protein, withthe VL and VH regions joined by the flexible linker (see e.g., Bird etal. (1988) Science 242:423-426; Huston et al. (1988) Proc. Natl. Acad.Sci. USA 85:5879-5883; McCafferty et al., Nature (1990) 348:552-554).

To express the antibodies, or antibody portions of the invention, DNAsencoding partial or full-length light and heavy chains, obtained asdescribed above, are inserted into expression vectors such that thegenes are operatively linked to transcriptional and translationalcontrol sequences. In this context, the term “operatively linked” isintended to mean that an antibody gene is ligated into a vector suchthat transcriptional and translational control sequences within thevector serve their intended function of regulating the transcription andtranslation of the antibody gene. The expression vector and expressioncontrol sequences are chosen to be compatible with the expression hostcell used. The antibody light chain gene and the antibody heavy chaingene can be inserted into separate vector or, more typically, both genesare inserted into the same expression vector. The antibody genes areinserted into the expression vector by standard methods (e.g., ligationof complementary restriction sites on the antibody gene fragment andvector, or blunt end ligation if no restriction sites are present).Prior to insertion of the D2E7 or D2E7-related light or heavy chainsequences, the expression vector may already carry antibody constantregion sequences. For example, one approach to converting the D2E7 orD2E7-related VH and VL sequences to full-length antibody genes is toinsert them into expression vectors already encoding heavy chainconstant and light chain constant regions, respectively, such that theVH segment is operatively linked to the CH segment(s) within the vectorand the VL segment is operatively linked to the CL segment within thevector. Additionally or alternatively, the recombinant expression vectorcan encode a signal peptide that facilitates secretion of the antibodychain from a host cell. The antibody chain gene can be cloned into thevector such that the signal peptide is linked in-frame to the aminoterminus of the antibody chain gene. The signal peptide can be animmunoglobulin signal peptide or a heterologous signal peptide (i.e., asignal peptide from a non-immunoglobulin protein).

In addition to the antibody chain genes, the recombinant expressionvectors of the invention carry regulatory sequences that control theexpression of the antibody chain genes in a host cell. The term“regulatory sequence” is intended to include promoters, enhancers andother expression control elements (e.g., polyadenylation signals) thatcontrol the transcription or translation of the antibody chain genes.Such regulatory sequences are described, for example, in Goeddel; GeneExpression Technology: Methods in Enzymology 185, Academic Press, SanDiego, Calif. (1990). It will be appreciated by those skilled in the artthat the design of the expression vector, including the selection ofregulatory sequences may depend on such factors as the choice of thehost cell to be transformed, the level of expression of protein desired,etc. Preferred regulatory sequences for mammalian host cell expressioninclude viral elements that direct high levels of protein expression inmammalian cells, such as promoters and/or enhancers derived fromcytomegalovirus (CMV) (such as the CMV promoter/enhancer), Simian Virus40 (SV40) (such as the SV40 promoter/enhancer), adenovirus, (e.g., theadenovirus major late promoter (AdMLP)) and polyoma. For furtherdescription of viral regulatory elements, and sequences thereof, seee.g., U.S. Pat. No. 5,168,062 by Stinski, U.S. Pat. No. 4,510,245 byBell et al. and U.S. Pat. No. 4,968,615 by Schaffner et al.

In addition to the antibody chain genes and regulatory sequences, therecombinant expression vectors of the invention may carry additionalsequences, such as sequences that regulate replication of the vector inhost cells (e.g., origins of replication) and selectable marker genes.The selectable marker gene facilitates selection of host cells intowhich the vector has been introduced (see e.g., U.S. Pat. Nos.4,399,216, 4,634,665 and 5,179,017, all by Axel et al.). For example,typically the selectable marker gene confers resistance to drugs, suchas G418, hygromycin or methotrexate, on a host cell into which thevector has been introduced. Preferred selectable marker genes includethe dihydrofolate reductase (DHFR) gene (for use in dhfr⁻ host cellswith methotrexate selection/amplification) and the neo gene (for G418selection).

For expression of the light and heavy chains, the expression vector(s)encoding the heavy and light chains is transfected into a host cell bystandard techniques. The various forms of the term “transfection” areintended to encompass a wide variety of techniques commonly used for theintroduction of exogenous DNA into a prokaryotic or eukaryotic hostcell, e.g., electroporation, calcium-phosphate precipitation,DEAE-dextran transfection and the like. Although it is theoreticallypossible to express the antibodies of the invention in eitherprokaryotic or eukaryotic host cells, expression of antibodies ineukaryotic cells, and most preferably mammalian host cells, is the mostpreferred because such eukaryotic cells, and in particular mammaliancells, are more likely than prokaryotic cells to assemble and secrete aproperly folded and immunologically active antibody. Prokaryoticexpression of antibody genes has been reported to be ineffective forproduction of high yields of active antibody (Boss and Wood (1985)Immunology Today 6:12-13).

Preferred mammalian host cells for expressing the recombinant antibodiesof the invention include Chinese Hamster Ovary (CHO cells) (includingdhfr− CHO cells, described in Urlaub and Chasin, (1980) Proc. Natl.Acad. Sci. USA 77:4216-4220, used with a DHFR selectable marker, e.g.,as described in Kaufman and Sharp (1982) Mol. Biol. 159:601-621), NSOmyeloma cells, COS cells and SP2 cells. When recombinant expressionvectors encoding antibody genes are introduced into mammalian hostcells, the antibodies are produced by culturing the host cells for aperiod of time sufficient to allow for expression of the antibody in thehost cells or, more preferably, secretion of the antibody into theculture medium in which the host cells are grown. Antibodies can berecovered from the culture medium using standard protein purificationmethods.

Host cells can also be used to produce portions of intact antibodies,such as Fab fragments or scFv molecules. It is understood thatvariations on the above procedure are within the scope of the presentinvention. For example, it may be desirable to transfect a host cellwith DNA encoding either the light chain or the heavy chain (but notboth) of an antibody of this invention. Recombinant DNA technology mayalso be used to remove some or all of the DNA encoding either or both ofthe light and heavy chains that is not necessary for binding to hTNFα.The molecules expressed from such truncated DNA molecules are alsoencompassed by the antibodies of the invention. In addition,bifunctional antibodies may be produced in which one heavy and one lightchain are an antibody of the invention and the other heavy and lightchain are specific for an antigen other than hTNFα by crosslinking anantibody of the invention to a second antibody by standard chemicalcrosslinking methods.

In a preferred system for recombinant expression of an antibody, orantigen-binding portion thereof, of the invention, a recombinantexpression vector encoding both the antibody heavy chain and theantibody light chain is introduced into dhfr-CHO cells by calciumphosphate-mediated transfection. Within the recombinant expressionvector, the antibody heavy and light chain genes are each operativelylinked to CMV enhancer/AdMLP promoter regulatory elements to drive highlevels of transcription of the genes. The recombinant expression vectoralso carries a DHFR gene, which allows for selection of CHO cells thathave been transfected with the vector using methotrexateselection/amplification. The selected transformant host cells areculture to allow for expression of the antibody heavy and light chainsand intact antibody is recovered from the culture medium. Standardmolecular biology techniques are used to prepare the recombinantexpression vector, transfect the host cells, select for transformants,culture the host cells and recover the antibody from the culture medium.

Recombinant human antibodies of the invention in addition to D2E7 or anantigen binding portion thereof, or D2E7-related antibodies disclosedherein can be isolated by screening of a recombinant combinatorialantibody library, preferably a scFv phage display library, preparedusing human VL and VH cDNAs prepared from mRNA derived from humanlymphocytes. Methodologies for preparing and screening such librariesare known in the art. In addition to commercially available kits forgenerating phage display libraries (e.g., the Pharmacia RecombinantPhage Antibody System, catalog no. 27-9400-01; and the StratageneSurfZAP™ phage display kit, catalog no. 240612), examples of methods andreagents particularly amenable for use in generating and screeningantibody display libraries can be found in, for example, Ladner et al.U.S. Pat. No. 5,223,409; Kang et al. PCT Publication No. WO 92/18619;Dower et al. PCT Publication No. WO 91/17271; Winter et al. PCTPublication No. WO 92/20791; Markland et al. PCT Publication No. WO92/15679; Breitling et al. PCT Publication No. WO 93/01288; McCaffertyet al. PCT Publication No. WO 92/01047; Garrard et al. PCT PublicationNo. WO 92/09690; Fuchs et al. (1991) Bio/Technology 9:1370-1372; Hay etal. (1992) Hum Antibod Hybridomas 3:81-85; Huse et al. (1989) Science246:1275-1281; McCafferty et al., Nature (1990) 348:552-554; Griffithset al. (1993) EMBO J 12:725-734; Hawkins et al. (1992) J Mol Biol226:889-896; Clackson et al. (1991) Nature 352:624-628; Gram et al.(1992) PNAS 89:3576-3580; Garrard et al. (1991) Bio/Technology9:1373-1377; Hoogenboom et al. (1991) Nuc Acid Res 19:4133-4137; andBarbas et al. (1991) PNAS 88:7978-7982.

In a preferred embodiment, to isolate human antibodies with highaffinity and a low off rate constant for hTNFα, a murine anti-hTNFαantibody having high affinity and a low off rate constant for hTNFα(e.g., MAK 195, the hybridoma for which has deposit number ECACC 87050801) is first used to select human heavy and light chain sequenceshaving similar binding activity toward hTNFα, using the epitopeimprinting methods described in Hoogenboom et al., PCT Publication No.WO 93/06213. The antibody libraries used in this method are preferablyscFv libraries prepared and screened as described in McCafferty et al.,PCT Publication No. WO 92/01047, McCafferty et al., Nature (1990)348:552-554; and Griffiths et al., (1993) EMBO J. 12:725-734. The scFvantibody libraries preferably are screened using recombinant human TNFαas the antigen.

Once initial human VL and VH segments are selected, “mix and match”experiments, in which different pairs of the initially selected VL andVH segments are screened for hTNFα binding, are performed to selectpreferred VL/VH pair combinations. Additionally, to further improve theaffinity and/or lower the off rate constant for hTNFα binding, the VLand VH segments of the preferred VL/VH pair(s) can be randomly mutated,preferably within the CDR3 region of VH and/or VL, in a processanalogous to the in vivo somatic mutation process responsible foraffinity maturation of antibodies during a natural immune response. Thisin vitro affinity maturation can be accomplished by amplifying VH and VLregions using PCR primers complimentary to the VH CDR3 or VL CDR3,respectively, which primers have been “spiked” with a random mixture ofthe four nucleotide bases at certain positions such that the resultantPCR products encode VH and VL segments into which random mutations havebeen introduced into the VH and/or VL CDR3 regions. These randomlymutated VH and VL segments can be rescreened for binding to hTNFα andsequences that exhibit high affinity and a low off rate for hTNFαbinding can be selected.

Following screening and isolation of an anti-hTNFα antibody of theinvention from a recombinant immunoglobulin display library, nucleicacid encoding the selected antibody can be recovered from the displaypackage (e.g., from the phage genome) and subcloned into otherexpression vectors by standard recombinant DNA techniques. If desired,the nucleic acid can be further manipulated to create other antibodyforms of the invention (e.g., linked to nucleic acid encoding additionalimmunoglobulin domains, such as additional constant regions). To expressa recombinant human antibody isolated by screening of a combinatoriallibrary, the DNA encoding the antibody is cloned into a recombinantexpression vector and introduced into a mammalian host cells, asdescribed in further detail in above.

Methods of isolating human antibodies with high affinity and a low offrate constant for hTNFα are also described in U.S. Pat. Nos. 6,090,382,6,258,562, and 6,509,015, each of which is incorporated by referenceherein.

The TNFα inhibitor may also be a TNF fusion protein, e.g., etanercept(Enbrel®, Amgen; described in WO 91/03553 and WO 09/406,476,incorporated by reference herein). In another embodiment, the TNFαinhibitor is a recombinant TNF binding protein (r-TBP-I) (Serono).

III. Treatment of Erosive Polyarthritis

The invention provides methods of treating erosive polyarthritiscomprising administering a TNFα inhibitor, including, for an example, aTNFα antibody, to a subject having erosive polyarthritis. The inventionalso describes methods for determining the efficacy of a TNFα inhibitorfor the treatment of erosive polyarthritis. Preferably, the TNFα ishuman TNFα and the subject is a human subject. In one embodiment, theTNFα inhibitor is adalimumab, also referred to as HUMIRA® or

D2E7. The use of TNFα inhibitors, including antibodies and antibodyportions, in the treatment of erosive polyarthritis, as well as methodsfor determining the efficacy of a TNFα inhibitor for the treatment oferosive polyarthritis, is discussed further below:

The term “polyarthritis” generally refers to inflammation, i.e.,swelling, tenderness, or warmth, at two or more joints of a subject.

As used herein, the term “erosive polyarthritis” refers to a subject whohas polyarthritis which is damaging to the joint.

The invention provides a method for treating erosive polyarthritiscomprising administration of a TNFα inhibitor, including, for anexample, a TNFα antibody. The invention also provides a method forinhibiting radiographic progression of joint disease associated witherosive polyarthritis. Methods for administering a TNFα antibody, or anantigen-binding portion thereof, for the treatment of erosivepolyarthritis are described in more detail below.

The invention provides a method for determining the efficacy of ananti-TNFα treatment for erosive polyarthritis. Measures for determiningsuch efficacy include tests which determine whether joint destruction orerosion is improved following treatment. For example, the Total ModifiedSharp Score (mTSS) of a subject may be used to determine improvements inerosive polyarthritis in the subject. The mTSS may also be used as anassay to determine the efficacy of a treatment for erosivepolyarthritis.

A Sharp score is an Xray measurement in changes in total joint damage asassessed by bone erosions and joint space narrowing (Sharp et al. (1971)Arthritis & Rheumatism 14:706; Sharp et al. (1985) Arthritis &Rheumatism 28:16). The mTSS is a measure of the extent and severity ofjoint damage based on evaluations of x-rays of patients' hands and feet.Joints are scored for both joint erosions and joint space narrowing. ThemTSS is the sum of the erosion score (ES) and the joint space narrowing(JSN) score and has, for example, a range of about 0 to about 398, where0=no damage. The ES is the sum of joint scores collected for 46 jointsand has a range, for example, of about 0 to about 230. The JSN is thesum of joint scores collected for 42 joints and has a range, forexample, of about 0 to about 168. A score of 0 would indicate no change.In one embodiment of the invention, the mTSS is determined by combiningthe joint space narrowing score having a range of about 0-192 and anerosion score having a range of about 0-378.

An improved or constant mTSS demonstrates that the TNFα inhibitor iseffective for treating erosive polyarthritis. In one embodiment,efficacy of a TNFα inhibitor for the treatment of erosive polyarthritisis evidenced by a lack of progression of joint disease, e.g., no changein Sharp score, in mTSS over time in a subject having erosivepolyarthritis. In another embodiment, efficacy of a TNFα inhibitor forthe treatment of erosive polyarthritis is evidenced by a decrease in theradiographic progression of joint disease, e.g., decrease in Sharpscore, in mTSS over time in a subject having erosive polyarthritis.

In one embodiment, the overall change in the mTSS between baseline and atime period following treatment with a TNFα inhibitor is between about0.9 and about −0.2. In another embodiment, the overall change in themTSS between baseline and a time period following treatment with a TNFαinhibitor is between about 0.5 and about −0.2. In still anotherembodiment, the overall change in the mTSS between baseline and a timeperiod following treatment with a TNFα inhibitor is between about 0.2and about −0.2.

It should be noted that ranges intermediate to the above recited scores,e.g., about −0.1 to about 0.3, are also intended to be part of thisinvention. For example, ranges of values using a combination of any ofthe above recited values as upper and/or lower limits are intended to beincluded.

While treatment of an inflammatory disease with an anti-inflammatoryagent may result in clinical improvements following treatment, there maystill be progressive joint damage resulting from erosive polyarthritis(Gladman et al. (1990) J Rheumatol 17:809; Hanly et al. (1988) Ann RheumDis 47:386). Thus, it is a feature of this invention to provide a methodfor treating erosive polyarthritis which may be associated with anotherdisorder. In a preferred embodiment, the invention includes treatment oferosive polyarthritis associated with a disorder in which TNFα activityis detrimental, including, but not limited to, rheumatoid arthritis(including juvenile rheumatoid arthritis), Crohn's disease, psoriasis,psoriatic arthritis, and ankylosing spondylitis. Erosive polyarthritismay also be associated with multicentric reticulohistiocytosis (MRH)(Santilli et al. (2002) Ann Rehum Dis 61: 485).

As used herein, the term “a disorder in which TNFα activity isdetrimental” is intended to include diseases and other disorders inwhich the presence of TNFα in a subject suffering from the disorder hasbeen shown to be or is suspected of being either responsible for thepathophysiology of the disorder or a factor that contributes to aworsening of the disorder. Accordingly, a disorder in which TNFαactivity is detrimental is a disorder in which inhibition of TNFαactivity is expected to alleviate the symptoms and/or progression of thedisorder. Such disorders may be evidenced, for example, by an increasein the concentration of TNFα in a biological fluid of a subjectsuffering from the disorder (e.g., an increase in the concentration ofTNFα in serum, plasma, synovial fluid, etc. of the subject), which canbe detected, for example, using an anti-TNFα antibody as describedabove. There are numerous examples of disorders in which TNFα activityis detrimental. The use of TNFα inhibitors for the treatment of erosivepolyarthritis associated with specific disorders is discussed furtherbelow:

A. Autoimmune Diseases

In one embodiment, the invention includes treatment of erosivepolyarthritis associated with an autoimmune disease. Erosivepolyarthritis may be found in patients suffering from autoimmunediseases, including forms of arthritis such as rheumatoid arthritis andjuvenile rheumatoid arthritis (Verloes (1998) Med Genet. 35:943). TNFαantibodies, such as adalimumab, may be used to treat autoimmunediseases, in particular those associated with erosive polyarthritis.Examples of such autoimmune conditions include rheumatoid arthritis,rheumatoid spondylitis, osteoarthritis and gouty arthritis, allergy,multiple sclerosis, autoimmune diabetes, autoimmune uveitis andnephrotic syndrome. Other examples of autoimmune conditions includemultisystem autoimmune diseases and autoimmune hearing loss. Otherexamples of autoimmune disease are described in U.S. application Ser.No. 10/622,932, incorporated by reference herein.

Juvenile Rheumatoid Arthritis

Tumor necrosis factor has been implicated in the pathophysiology ofjuvenile arthritis, including juvenile rheumatoid arthritis (Grom et al.(1996) Arthritis Rheum. 39:1703; Mangge et al. (1995) Arthritis Rheum.8:211). In one embodiment, the TNFα antibody of the invention is used totreat juvenile rheumatoid arthritis.

The term “juvenile rheumatoid arthritis” or “JRA” as used herein refersto a chronic, inflammatory disease which occurs before age 16 that maycause joint or connective tissue damage. JRA is also referred to asjuvenile chronic polyarthritis and Still's disease.

JRA causes joint inflammation and stiffness for more than 6 weeks in achild of 16 years of age or less. Inflammation causes redness, swelling,warmth, and soreness in the joints. Any joint can be affected andinflammation may limit the mobility of affected joints. One type of JRAcan also affect the internal organs.

JRA is often classified into three types by the number of jointsinvolved, the symptoms, and the presence or absence of certainantibodies found by a blood test. These classifications help thephysician determine how the disease will progress and whether theinternal organs or skin is affected. The classifications of JRA includethe following:

a. Pauciarticular JRA, wherein the patient has four or fewer joints areaffected. Pauciarticular is the most common form of JRA, and typicallyaffects large joints, such as the knees.

b. Polyarticular HRA, wherein five or more joints are affected. Thesmall joints, such as those in the hands and feet, are most commonlyinvolved, but the disease may also affect large joints.

c. Systemic JRA is characterized by joint swelling, fever, a light skinrash, and may also affect internal organs such as the heart, liver,spleen, and lymph nodes. Systemic JRA is also referred to as it Still'sdisease. A small percentage of these children develop arthritis in manyjoints and can have severe arthritis that continues into adulthood.

B. Spondyloarthropathies

In one embodiment, the invention includes treatment of erosivepolyarthritis associated with a spondylarthopathy. Erosive polyarthritismay be found in patients suffering from inflammatory diseases, such asspondyloarthopathies, associated with detrimental TNFα activity (seee.g., Moeller et al. (1990) Cytokine 2:162; U.S. Pat. No. 5,231,024;European Patent Publication No. 260 610).

As used herein, the term “spondyloarthropathy” or“spondyloarthropathies” is used to refer to any one of several diseasesaffecting the joints of the spine, wherein such diseases share commonclinical, radiological, and histological features. A number ofspondyloarthropathies share genetic characteristics, i.e. they areassociated with the HLA-B27 allele. In one embodiment, the termspondyloarthropathy is used to refer to any one of several diseasesaffecting the joints of the spine, excluding ankylosing spondylitis,wherein such diseases share common clinical, radiological, andhistological features. Examples of spondyloarthropathies includeankylosing spondylitis, psoriatic arthritis/spondylitis, enteropathicarthritis, reactive arthritis or Reiter's syndrome, and undifferentiatedspondyloarthropathies. Examples of animal models used to studyspondyloarthropathies include ank/ank transgenic mice, HLA-B27transgenic rats (see Taurog et al. (1998) The Spondylarthritides.Oxford:Oxford University Press).

Examples of subjects who are at risk of having spondyloarthropathiesinclude humans suffering from arthritis. Spondyloarthropathies can beassociated with forms of arthritis, including rheumatoid arthritis. Inone embodiment of the invention, a TNFα inhibitor is used to treat asubject who suffers from a spondyloarthropathy associated with erosivepolyarthritis. Examples of spondyloarthropathies which can be treatedwith a TNFα inhibitor are described below:

Ankylosing Spondylitis (AS)

In one embodiment, the invention includes treatment of erosivepolyarthritis associated with ankylosing spondylitis using a TNFαantibody, or antigen-binding portion thereof. Tumor necrosis factor hasbeen implicated in the pathophysiology of ankylosing spondylitis (seeVerjans et al. (1991) Arthritis Rheum. 34:486; Verjans et al. (1994)Clin Exp Immunol. 97:45; Kaijtzel et al. (1999) Hum Immunol. 60:140).Ankylosing spondylitis (AS) is an inflammatory disorder involvinginflammation of one or more vertebrae. AS is a chronic inflammatorydisease that affects the axial skeleton and/or peripheral joints,including joints between the vertebrae of the spine and sacroiliacjoints and the joints between the spine and the pelvis. AS caneventually cause the affected vertebrae to fuse or grow together.Spondyarthropathies, including AS, can be associated with psoriaticarthritis (PsA) and/or inflammatory bowel disease (IBD), includingulcerative colitis and Crohn's disease.

Early manifestations of AS can be determined by radiographic tests,including CT scans and MRI scans. Early manifestations of AS ofteninclude scroiliitis and changes in the sacroliac joints as evidenced bythe blurring of the cortical margins of the subchrondral bone, followedby erosions and sclerosis. Fatigue has also been noted as a commonsymptom of AS (Duffy et al. (2002) ACR 66th Annual Scientific MeetingAbstract).

Psoriatic Arthritis

In one embodiment, the invention includes treatment of erosivepolyarthritis associated with psoriatic arthritis using a TNFα antibody,or antigen-binding portion thereof. Tumor necrosis factor has beenimplicated in the pathophysiology of psoriatic arthritis (PsA) (Partschet al. (1998) Ann Rheum Dis. 57:691; Ritchlin et al. (1998) J Rheumatol.25:1544). As referred to herein, psoriatic TNFα has been implicated inactivating tissue inflammation and causing joint destruction inrheumatoid arthritis (see e.g., Moeller, A., et al. (1990) Cytokine2:162-169; U.S. Pat. No. 5,231,024 to Moeller et al.; European PatentPublication No. 260 610 B1 by Moeller, A.; Tracey and Cerami, supra;Arend, W. P. and Dayer, J-M. (1995) Arth. Rheum. 38:151-160; Fava, R.A., et al. (1993) Clin. Exp. Immunol. 94:261-266). TNFα also has beenimplicated in promoting the death of islet cells and in mediatinginsulin resistance in diabetes (see e.g., Tracey and Cerami, supra; PCTPublication No. WO 94/08609). TNFα also has been implicated in mediatingcytotoxicity to oligodendrocytes and induction of inflammatory plaquesin multiple sclerosis (see e.g., Tracey and Cerami, supra). Chimeric andhumanized murine anti-hTNFα antibodies have undergone clinical testingfor treatment of rheumatoid arthritis (see e.g., Elliott, M. J., et al.(1994) Lancet 344:1125-1127; Elliot, M. J., et al. (1994) Lancet344:1105-1110; Rankin, E. C., et al. (1995) Br. J. Rheumatol.34:334-342).

Psoriatic arthritis refers to chronic inflammatory arthritis which isassociated with psoriasis, a common chronic skin condition that causesred patches on the body. About 1 in 20 individuals with psoriasis willdevelop arthritis along with the skin condition, and in about 75% ofcases, psoriasis precedes the arthritis. PsA exhibits itself in avariety of ways, ranging from mild to severe arthritis, wherein thearthritis usually affects the fingers and the spine. When the spine isaffected, the symptoms are similar to those of ankylosing spondylitis,as described above. A TNFα antibody, or antigen-binding fragmentthereof, can be used for treatment of erosive polyarthritis associatedwith PsA.

PsA is sometimes associated with arthritis mutilans. Arthritis mutilansrefers to a disorder which is characterized by excessive bone erosionresulting in a gross, erosive deformity which mutilates the joint.

Characteristic radiographic features of PsA include joint erosions,joint space narrowing, bony proliferation including periarticular andshaft periostitis, osteolysis including “pencil in cup” deformity andacro-osteolysis, ankylosis, spur formation, and spondylitis (Wassenberget al. (2001) Z Rheumatol 60:156). Unlike rheumatoid arthritis (RA),joint involvement in PsA is often asymmetrical and may beoligoarticular; osteoporosis is atypical. Although erosive changes inearly PsA are marginal as in RA, they become irregular and ill definedwith disease progression because of periosteal bone formation adjacentto the erosions. In severe cases, erosive changes may progress todevelopment of pencil in cup deformity or gross osteolysis (Gold et al.(1988) Radiol Clin North Am 26:1195; Resnick et al. (1977)) J Can AssocRadiol 28:187). Asymmetrical erosions may be visible radiographically inthe carpus and in the metacarpophalangeal (MCP), proximalinterphalangeal (PIP), and distal interphalangeal (DIP) joints of thehands, but the DIP joints are often the first to be affected.Abnormalities are seen in the phalangeal tufts and at the sites ofattachments of tendons and ligaments to the bone. The presence of DIPerosive changes may provide both sensitive and specific radiographicfindings to support the diagnosis of PsA. Also, the hands tend to beinvolved much more frequently than the feet with a ratio of nearly 2:1.

Other examples of spondyloarthropathies are described in U.S.application Ser. No. 10/622,932, incorporated by reference herein.

C. Skin and Nail Disorders

In one embodiment, the invention includes treatment of erosivepolyarthritis associated with skin and nail disorders. As used herein,the term “skin and nail disorder in which TNFα activity is detrimental”is intended to include skin and/or nail disorders and other disorders inwhich the presence of TNFα in a subject suffering from the disorder hasbeen shown to be or is suspected of being either responsible for thepathophysiology of the disorder or a factor that contributes to aworsening of the disorder, e.g., psoriasis. Accordingly, skin and naildisorders in which TNFα activity is detrimental are disorders in whichinhibition of TNFα activity is expected to alleviate the symptoms and/orprogression of the disorder. The use of the antibodies, antibodyportions, and other TNFα inhibitors for the treatment of specific skinand nail disorders is discussed further below. In certain embodiments,the antibody, antibody portion, or other TNFα inhibitor of the inventionis administered to the subject in combination with another therapeuticagent, as described below. In one embodiment, a TNFα antibody isadministered to the subject in combination with another therapeuticagent for the treatment of erosive polyarthritis associated withpsoriasis and the treatment of psoriasis associated with arthritis.

Psoriasis

Tumor necrosis factor has been implicated in the pathophysiology ofpsoriasis (Takematsu et al. (1989) Arch Dermatol Res. 281:398; Victorand Gottlieb (2002) J Drugs Dermatol. 1(3):264). Psoriasis is describedas a skin inflammation (irritation and redness) characterized byfrequent episodes of redness, itching, and thick, dry, silvery scales onthe skin. In particular, lesions are formed which involve primary andsecondary alterations in epidermal proliferation, inflammatory responsesof the skin, and an expression of regulatory molecules such aslymphokines and inflammatory factors. Psoriatic skin is morphologicallycharacterized by an increased turnover of epidermal cells, thickenedepidermis, abnormal keratinization, inflammatory cell infiltrates intothe epidermis and polymorphonuclear leukocyte and lymphocyteinfiltration into the epidermis layer resulting in an increase in thebasal cell cycle. Psoriasis often involves the nails, which frequentlyexhibit pitting, separation of the nail, thickening, and discoloration.Psoriasis is often associated with other inflammatory disorders, forexample arthritis, including rheumatoid arthritis, inflammatory boweldisease (IBD), and Crohn's disease.

Evidence of psoriasis is most commonly seen on the trunk, elbows, knees,scalp, skin folds, or fingernails, but it may affect any or all parts ofthe skin. Normally, it takes about a month for new skin cells to move upfrom the lower layers to the surface. In psoriasis, this process takesonly a few days, resulting in a build-up of dead skin cells andformation of thick scales. Symptoms of psoriasis include: skin patches,that are dry or red, covered with silvery scales, raised patches ofskin, accompanied by red borders, that may crack and become painful, andthat are usually lovated on the elbows, knees, trunk, scalp, and hands;skin lesions, including pustules, cracking of the skin, and skinredness; joint pain or aching which may be associated with of arthritis,e.g., psoriatic arthritis.

Treatment for psoriasis often includes a topical corticosteroids,vitamin D analogs, and topical or oral retinoids, or combinationsthereof. In one embodiment, the TNFα inhibitor of the invention isadministered in combination with or the presence of one of these commontreatments. Additional therapeutic agents which can also be combinedwith the TNFα inhibitor for treatment of psoriasis are described in moredetail below.

The diagnosis of psoriasis is usually based on the appearance of theskin. Additionally a skin biopsy, or scraping and culture of skinpatches may be needed to rule out other skin disorders. An x-ray may beused to check for psoriatic arthritis if joint pain is present andpersistent.

In one embodiment of the invention, a TNFα inhibitor is used to treatpsoriasis, including chronic plaque psoriasis, guttate psoriasis,inverse psoriasis, pustular psoriasis, pemphigus vulgaris, erythrodermicpsoriasis, psoriasis associated with inflammatory bowel disease (IBD),and psoriasis associated with rheumatoid arthritis (RA). Specific typesof psoriasis included in the treatment methods of the invention includechronic plaque psoriasis, guttate psoriasis, inverse psoriasis, andpustular psoriasis. Other examples of psoriasis and other types of skinand nail disorders are described in U.S. application Ser. No. 10/622,932(U.S. Pat. Publ. 2004/0126372), incorporated by reference herein.

Methods for determining the efficacy of a TNFα inhibitor for thetreatment of erosive polyarthritis in association a disorder in whichTNFα activity is detrimental include any assay which measures the degreeof joint destruction, including joint space narrowing and/or jointerosion. In one embodiment, joint destruction is measured usingradiography. Such assays may be used to examine the efficacy of the TNFαinhibitor by determining whether an improvement occurs in a subject orpatient population treated with the TNFα inhibitor. Generally,improvements are determined by comparing a baseline score determinedprior to treatment, and a score determined at a time following treatmentwith the TNFα inhibitor.

Additional improvements in arthritic conditions, such as Ra, PsA, andJRA, may be determined by measuring the ACR response. ACR thresholds,e.g., ACR20, ACR50, ACR70, may be used for defining improvement in RAand PsA, and indicate the percentage improvement in seven diseaseactivity measures. Criteria include percentage improvement in tender andswollen joint count and improvement of at least 3 of the followingcriteria: patient pain assessment, patient global assessment, physicianglobal assessment, patient self-assessed disability, or laboratorymeasures of disease activity (i.e., erythrocyte sedimentation rate orC-reactive protein level) (Felson et al. (1993) Arthritis Rheum.36(6):729).

Other assays used to determine improvement for a given therapy for thetreatment of RA, JRA, and PsA, include the EULAR response, DAS score,FACIT-F, HAQ score, and SJC and/or TJC counts.

The EULAR criteria uses a DAS for defining response. Response is definedas both: (a) change in disease activity from baseline and (b) the levelof disease activity reached during follow-up. Criteria used to defineDAS include: Ritchie articular index, swollen joint count (44-jointcount), erythrocyte sedimentation rate, and Health AssessmentQuestionnaire. A modified version of the DAS criteria, DAS28, uses a28-joint count for swollen and tender joints. Response is defined as acombination of a significant change from baseline and the level ofdisease activity attained. Good response is defined as a significantdecrease in DAS (>1.2) and a low level of disease activity (< or =2.4).Non-response is defined as a decrease < or =0.6, or a decrease >0.6 and< or =1.2 with an attained DAS>3.7. Any other scores are regarded asmoderate responses.

The DAS is a score is based on the Ritchie articular index, a 44 swollenjoint count, ESR, and a general health assessment on a VAS. Range variesfrom 1 to 9. Serial measurements of the DAS and DAS28 are strongpredictors of physical disability and radiological progression, and bothindices are sensitive discriminators between patients with high and lowdisease activity and between active and placebo treated patient groups.

FACIT-F (Functional Assessment of Chronic Illness Therapy-Fatigue) is avalidated questionnaire designed to measure patients' assessment offatigue-related factors in chronic illness (see Cella and Webster (1997)Oncology (Huntingt). 11:232 and Lai et al. (2003) Qual Life Res.12(5):485).

The Health Assessment Questionnaire (HAQ) is a validated questionnairedesigned to assess patients' ability to perform activities of dailyliving, particularly in adult arthritics. Instrument consists of the HAQDisability Index (20 items), Pain Scale (1 item), and Global HealthStatus (1 item) that measure disability/physical functioning and qualityof life (Fries et al. (1982) J Rheumatol. 9(5):789).

Swollen and tender joints (SJC and TJC) are the most characteristicfeatures of RA, and disease severity is directly related to the numberof swollen and tender joints. Counting swollen and tender joints is akey component of the clinical assessment of RA.

Improvements in PsA and psoriasis may also be determined using the PASIresponse, DLQI, and the BSA score. DLQI (Dermatology Life Quality Index)is a health-related quality of life measure widely used for a variety ofdermatological diseases, including PsA and psoriasis. The body SurfaceArea (BSA) score provides a measurement of surface area based on heightand weight and expressed in m². The PASI (Psoriasis Area and SeverityIndex) is a composite measure of the erythema, induration, desquamationand body surface area that is affected by psoriasis for a particularpatient. Patients are evaluated for head, trunk, upper and lower limbinvolvement. Scores range from 0 (clear) to 72 (maximum severity).

Improvements in treatment for PsA may also be measured using the PsARC(Psoriatic Arthritis Response Criteria), which provides a clinicalmeasure of the change in tender and swollen joint scores, along with aseries of global assessments of disease activity.

Improvements in AS may be measured by using any number of instruments toevaluate various AS symptoms. Some of the commonly used scales includethe Assessment in Ankylosing Spondylitis (ASAS), the Bath AnkylosingSpondylitis Disease Activity Index (BASDAI) (Garrett et al. (1994) JRheumatol 21:2286), the Bath Ankylosing Spondylitis Metrology Index(BASMI) (Jenkinson et al. (1994) J Rheumatol 21:1694), and the BathAnkylosing Spondylitis Functional Index (BASFI) (Calin et al. (1994) JRheumatol 21:2281). These indices can be used to monitor a patient overtime and to determine improvement. Additional description measurementsfor assessing improvements in AS are described in U.S. application Ser.No. 10/622,932, incorporated by reference herein.

IV. Pharmaceutical Compositions and Pharmaceutical Administration

A. Compositions and Administration

Antibodies, antibody-portions, and other TNFα inhibitors for use in themethods of the invention, may be incorporated into pharmaceuticalcompositions suitable for administration to a subject having erosivepolyarthritis. Typically, the pharmaceutical composition comprises anantibody, antibody portion, or other TNFα inhibitor of the invention anda pharmaceutically acceptable carrier. As used herein, “pharmaceuticallyacceptable carrier” includes any and all solvents, dispersion media,coatings, antibacterial and antifungal agents, isotonic and absorptiondelaying agents, and the like that are physiologically compatible.Examples of pharmaceutically acceptable carriers include one or more ofwater, saline, phosphate buffered saline, dextrose, glycerol, ethanoland the like, as well as combinations thereof. In many cases, it ispreferable to include isotonic agents, for example, sugars, polyalcoholssuch as mannitol, sorbitol, or sodium chloride in the composition.Pharmaceutically acceptable carriers may further comprise minor amountsof auxiliary substances such as wetting or emulsifying agents,preservatives or buffers, which enhance the shelf life or effectivenessof the antibody, antibody portion, or other TNFα inhibitor.

The compositions for use in the methods of the invention may be in avariety of forms. These include, for example, liquid, semi-solid andsolid dosage forms, such as liquid solutions (e.g., injectable andinfusible solutions), dispersions or suspensions, tablets, pills,powders, liposomes and suppositories. The preferred form depends on theintended mode of administration and therapeutic application. Typicalpreferred compositions are in the form of injectable or infusiblesolutions, such as compositions similar to those used for passiveimmunization of humans with other antibodies or other TNFα inhibitors.The preferred mode of administration is parenteral (e.g., intravenous,subcutaneous, intraperitoneal, intramuscular). In one embodiment, theantibody or other TNFα inhibitor is administered by intravenous infusionor injection. In another embodiment, the antibody or other TNFαinhibitor is administered by intramuscular or subcutaneous injection.

Therapeutic compositions typically must be sterile and stable under theconditions of manufacture and storage. The composition can be formulatedas a solution, microemulsion, dispersion, liposome, or other orderedstructure suitable to high drug concentration. Sterile injectablesolutions can be prepared by incorporating the active compound (i.e.,antibody, antibody portion, or other TNFα inhibitor) in the requiredamount in an appropriate solvent with one or a combination ofingredients enumerated above, as required, followed by filteredsterilization. Generally, dispersions are prepared by incorporating theactive compound into a sterile vehicle that contains a basic dispersionmedium and the required other ingredients from those enumerated above.In the case of sterile powders for the preparation of sterile injectablesolutions, the preferred methods of preparation are vacuum drying andfreeze-drying that yields a powder of the active ingredient plus anyadditional desired ingredient from a previously sterile-filteredsolution thereof. The proper fluidity of a solution can be maintained,for example, by the use of a coating such as lecithin, by themaintenance of the required particle size in the case of dispersion andby the use of surfactants. Prolonged absorption of injectablecompositions can be brought about by including in the composition anagent that delays absorption, for example, monostearate salts andgelatin.

Supplementary active compounds can also be incorporated into thecompositions. In certain embodiments, an antibody or antibody portionfor use in the methods of the invention is coformulated with and/orcoadministered with one or more additional therapeutic agents, includingan erosive polyarthritis inhibitor or antagonist. For example, ananti-hTNFα antibody or antibody portion of the invention may becoformulated and/or coadministered with one or more additionalantibodies that bind other targets associated with erosive polyarthritis(e.g., antibodies that bind other cytokines or that bind cell surfacemolecules), one or more cytokines, soluble TNFα receptor (see e.g., PCTPublication No. WO 94/06476) and/or one or more chemical agents thatinhibit hTNFα production or activity (such as cyclohexane-ylidenederivatives as described in PCT Publication No. WO 93/19751) or anycombination thereof. Furthermore, one or more antibodies of theinvention may be used in combination with two or more of the foregoingtherapeutic agents. Such combination therapies may advantageouslyutilize lower dosages of the administered therapeutic agents, thusavoiding possible side effects, complications or low level of responseby the patient associated with the various monotherapies.

In one embodiment, the invention includes pharmaceutical compositionscomprising an effective amount of a TNFα inhibitor and apharmaceutically acceptable carrier, wherein the effective amount of theTNFα inhibitor may be effective to treat erosive polyarthritis. In oneembodiment, the antibody or antibody portion for use in the methods ofthe invention is incorporated into a pharmaceutical formulation asdescribed in PCT/IB03/04502 and U.S. application Ser. No. 10/222,140,incorporated by reference herein. This formulation includes aconcentration 50 mg/ml of the antibody D2E7, wherein one pre-filledsyringe contains 40 mg of antibody for subcutaneous injection. Inanother embodiment, the formulation of the invention includes D2E7.

The antibodies, antibody-portions, and other TNFα inhibitors of thepresent invention can be administered by a variety of methods known inthe art, although for many therapeutic applications, the preferredroute/mode of administration is subcutaneous injection. In anotherembodiment, administration is via intravenous injection or infusion. Aswill be appreciated by the skilled artisan, the route and/or mode ofadministration will vary depending upon the desired results. In certainembodiments, the active compound may be prepared with a carrier thatwill protect the compound against rapid release, such as a controlledrelease formulation, including implants, transdermal patches, andmicroencapsulated delivery systems. Biodegradable, biocompatiblepolymers can be used, such as ethylene vinyl acetate, polyanhydrides,polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Manymethods for the preparation of such formulations are patented orgenerally known to those skilled in the art. See, e.g., Sustained andControlled Release Drug Delivery Systems, Robinson, ed., Dekker, Inc.,New York, 1978.

TNFα antibodies may also be administered in the form of protein crystalformulations which include a combination of protein crystalsencapsulated within a polymeric carrier to form coated particles. Thecoated particles of the protein crystal formulation may have a sphericalmorphology and be microspheres of up to 500 micro meters in diameter orthey may have some other morphology and be microparticulates. Theenhanced concentration of protein crystals allows the antibody of theinvention to be delivered subcutaneously. In one embodiment, the TNFαantibodies of the invention are delivered via a protein delivery system,wherein one or more of a protein crystal formulation or composition, isadministered to a subject with a TNFα-related disorder. Compositions andmethods of preparing stabilized formulations of whole antibody crystalsor antibody fragment crystals are also described in WO 02/072636, whichis incorporated by reference herein. In one embodiment, a formulationcomprising the crystallized antibody fragments described inPCT/IB03/04502 and U.S. application Ser. No. 10/222,140, incorporated byreference herein, are used to treat erosive polyarthritis usingtreatment methods of the invention.

In certain embodiments, an antibody, antibody portion, or other TNFαinhibitor of the invention may be orally administered, for example, withan inert diluent or an assimilable edible carrier. The compound (andother ingredients, if desired) may also be enclosed in a hard or softshell gelatin capsule, compressed into tablets, or incorporated directlyinto the subject's diet. For oral therapeutic administration, thecompounds may be incorporated with excipients and used in the form ofingestible tablets, buccal tablets, troches, capsules, elixirs,suspensions, syrups, wafers, and the like. To administer a compound ofthe invention by other than parenteral administration, it may benecessary to coat the compound with, or co-administer the compound with,a material to prevent its inactivation.

The pharmaceutical compositions of the invention may include a“therapeutically effective amount” or a “prophylactically effectiveamount” of an antibody or antibody portion of the invention. A“therapeutically effective amount” refers to an amount effective, atdosages and for periods of time necessary, to achieve the desiredtherapeutic result. A therapeutically effective amount of the antibody,antibody portion, or other TNFα inhibitor may vary according to factorssuch as the disease state, age, sex, and weight of the individual, andthe ability of the antibody, antibody portion, other TNFα inhibitor toelicit a desired response in the individual. A therapeutically effectiveamount is also one in which any toxic or detrimental effects of theantibody, antibody portion, or other TNFα inhibitor are outweighed bythe therapeutically beneficial effects. A “prophylactically effectiveamount” refers to an amount effective, at dosages and for periods oftime necessary, to achieve the desired prophylactic result. Typically,since a prophylactic dose is used in subjects prior to or at an earlierstage of disease, the prophylactically effective amount will be lessthan the therapeutically effective amount.

Dosage regimens may be adjusted to provide the optimum desired response(e.g., a therapeutic or prophylactic response). For example, a singlebolus may be administered, several divided doses may be administeredover time or the dose may be proportionally reduced or increased asindicated by the exigencies of the therapeutic situation. It isespecially advantageous to formulate parenteral compositions in dosageunit form for ease of administration and uniformity of dosage. Dosageunit form as used herein refers to physically discrete units suited asunitary dosages for the mammalian subjects to be treated; each unitcontaining a predetermined quantity of active compound calculated toproduce the desired therapeutic effect in association with the requiredpharmaceutical carrier. The specification for the dosage unit forms ofthe invention are dictated by and directly dependent on (a) the uniquecharacteristics of the active compound and the particular therapeutic orprophylactic effect to be achieved, and (b) the limitations inherent inthe art of compounding such an active compound for the treatment ofsensitivity in individuals.

In one embodiment, the invention provides a single dose method fortreating erosive polyarthritis, comprising administering to a subject inneed thereof a single dose of a TNFα inhibitor, such as a humanantibody. In one embodiment, the TNFα inhibitor is the anti-TNFαantibody adalimumab. The single dose of TNFα inhibitor can be anytherapeutically or prophylactically effective amount. In one embodiment,a subject is administered either a 20 mg, a 40 mg, or an 80 mg singledose of adalimumab (also referred to as D2E7). The single dose may beadministered through any route, including, for example, subcutaneousadministration. Biweekly dosing regimens can be used to treat erosivepolyarthritis and are further described in U.S. application Ser. No.10/163,657. Multiple variable dose methods of treatment or preventioncan also be used to treat erosive polyarthritis, and are furtherdescribed in PCT appln. no. PCT/US05/012007.

It is to be noted that dosage values may vary with the type and severityof the condition to be alleviated. It is to be further understood thatfor any particular subject, specific dosage regimens should be adjustedover time according to the individual need and the professional judgmentof the person administering or supervising the administration of thecompositions, and that dosage ranges set forth herein are exemplary onlyand are not intended to limit the scope or practice of the claimedcomposition. It should also be noted that the invention pertains tomethods of treatment of erosive polyarthritis including acute managementand chronic management of the disease.

The invention also pertains to packaged pharmaceutical compositions orkits for administering the anti-TNF antibodies of the invention for thetreatment of erosive polyarthritis. In one embodiment of the invention,the kit comprises a TNFα inhibitor, such as an antibody, a secondpharmaceutical composition comprising an additional therapeutic agent,and instructions for administration for treatment of erosivepolyarthritis. The instructions may describe how, e.g., subcutaneously,and when, e.g., at week 0 and week 2, the different doses of TNFαinhibitor and/or the additional therapeutic agent shall be administeredto a subject for treatment.

Another aspect of the invention pertains to kits containing apharmaceutical composition comprising an anti-TNFα antibody and apharmaceutically acceptable carrier and one or more pharmaceuticalcompositions each comprising a drug useful for treating erosivepolyarthritis and a pharmaceutically acceptable carrier. Alternatively,the kit comprises a single pharmaceutical composition comprising ananti-TNFα antibody, one or more drugs useful for treating erosivepolyarthritis and a pharmaceutically acceptable carrier. The kitscontain instructions for dosing of the pharmaceutical compositions forthe treatment of erosive polyarthritis.

The package or kit alternatively can contain the TNFα inhibitor and itcan be promoted for use, either within the package or throughaccompanying information, for the uses or treatment of the disordersdescribed herein. The packaged pharmaceuticals or kits further caninclude a second agent (as described herein) packaged with or copromotedwith instructions for using the second agent with a first agent (asdescribed herein).

B. Additional Therapeutic Agents

The invention also describes methods of treating erosive polyarthritiscomprising administering a TNFα inhibitor in combination with anadditional therapeutic agent. The invention also pertains topharmaceutical compositions and methods of use thereof for the treatmentof erosive polyarthritis in combination with an additional therapeuticagent. The pharmaceutical compositions comprise a first agent thattreats erosive polyarthritis. The pharmaceutical composition also maycomprise a second agent that is an active pharmaceutical ingredient;that is, the second agent is therapeutic and its function is beyond thatof an inactive ingredient, such as a pharmaceutical carrier,preservative, diluent, or buffer. In one embodiment, the second agentmay be useful in treating or preventing erosive polyarthritis. Inanother embodiment, the second agent may diminish or treat at least onesymptom(s) associated with the disorder which is associated with erosivepolyarthritis, e.g., psoriatic arthritis. In yet another embodiment, theadditional agent is useful for the treatment of both erosivepolyarthritis and the additional disorder. The first and second agentsmay exert their biological effects by similar or unrelated mechanisms ofaction; or either one or both of the first and second agents may exerttheir biological effects by a multiplicity of mechanisms of action. Apharmaceutical composition may also comprise a third compound, or evenmore yet, wherein the third (and fourth, etc.) compound has the samecharacteristics of a second agent.

It should be understood that the pharmaceutical compositions describedherein may have the first and second, third, or additional agents in thesame pharmaceutically acceptable carrier or in a differentpharmaceutically acceptable carrier for each described embodiment. Itfurther should be understood that the first, second, third andadditional agent may be administered simultaneously or sequentiallywithin described embodiments. Alternatively, a first and second agentmay be administered simultaneously, and a third or additional agent maybe administered before or after the first two agents.

The combination of agents used within the methods and pharmaceuticalcompositions described herein may have a therapeutic additive orsynergistic effect on the condition(s) or disease(s) targeted fortreatment. The combination of agents used within the methods orpharmaceutical compositions described herein also may reduce adetrimental effect associated with at least one of the agents whenadministered alone or without the other agent(s) of the particularpharmaceutical composition. For example, the toxicity of side effects ofone agent may be attenuated by another agent of the composition, thusallowing a higher dosage, improving patient compliance, and improvingtherapeutic outcome. The additive or synergistic effects, benefits, andadvantages of the compositions apply to classes of therapeutic agents,either structural or functional classes, or to individual compoundsthemselves.

Supplementary active compounds can also be incorporated into thecompositions. In certain embodiments, an antibody or antibody portion ofthe invention is coformulated with and/or coadministered with one ormore additional therapeutic agents that are useful for treating erosivepolyarthritis. For example, an anti-hTNFα antibody, antibody portion, orother TNFα inhibitor of the invention may be coformulated and/orcoadministered with one or more additional antibodies that bind othertargets (e.g., antibodies that bind other cytokines or that bind cellsurface molecules), one or more cytokines, soluble TNFα receptor (seee.g., PCT Publication No. WO 94/06476) and/or one or more chemicalagents that inhibit hTNFα production or activity (such ascyclohexane-ylidene derivatives as described in PCT Publication No. WO93/19751). Furthermore, one or more antibodies or other TNFα inhibitorsof the invention may be used in combination with two or more of theforegoing therapeutic agents. Such combination therapies mayadvantageously utilize lower dosages of the administered therapeuticagents, thus avoiding possible toxicities or complications associatedwith the various monotherapies.

TNFα inhibitors, e.g., a TNFα antibody, or antigen-binding portionthereof, described herein may be used in combination with additionaltherapeutic agents for the treatment erosive polyarthritis. Preferablythe other drug is a Disease Modifying Anti-Rheumatic Drug (DMARD) or aNonsteroidal Antiinflammatory Drug (NSAID) or a steroid or anycombination thereof. Preferred examples of a DMARD arehydroxychloroquine, leflunomide, methotrexate, parenteral gold, oralgold and sulfasalazine.

Nonlimiting additional agents which can also be used in combination witha TNFα inhibitor, e.g., a TNFα antibody, or antigen-binding portionthereof, to treat erosive polyarthritis include, but are not limited to,the following: non-steroidal anti-inflammatory drug(s) (NSAIDs);cytokine suppressive anti-inflammatory drug(s) (CSAIDs);CDP-571/BAY-10-3356 (humanized anti-TNFα antibody; Celltech/Bayer);cA2/infliximab (chimeric anti-TNFα antibody; Centocor); 75kdTNFR-IgG/etanercept (75 kD TNF receptor-IgG fusion protein; Immunex;see e.g., Arthritis & Rheumatism (1994) Vol. 37, 5295; J. Invest. Med.(1996) Vol. 44, 235A); 55 kdTNF-IgG (55 kD TNF receptor-IgG fusionprotein; Hoffmann-LaRoche); IDEC-CE9.1/SB 210396 (non-depletingprimatized anti-CD4 antibody; IDEC/SmithKline; see e.g., Arthritis &Rheumatism (1995) Vol. 38, S185); DAB 486-IL-2 and/or DAB 389-IL-2 (IL-2fusion proteins; Seragen; see e.g., Arthritis & Rheumatism (1993) Vol.36, 1223); Anti-Tac (humanized anti-IL-2Ra; Protein Design Labs/Roche);IL-4 (anti-inflammatory cytokine; DNAX/Schering); IL-10 (SCH 52000;recombinant IL-10, anti-inflammatory cytokine; DNAX/Schering); IL-4;IL-10 and/or IL-4 agonists (e.g., agonist antibodies); IL-1RA (IL-1receptor antagonist; Synergen/Amgen); anakinra (Kineret®/Amgen):TNF-bp/s-TNF (soluble TNF binding protein; see e.g., Arthritis &Rheumatism (1996) Vol. 39, No. 9 (supplement), S284; Amer. J.Physiol.—Heart and Circulatory Physiology (1995) Vol. 268, pp. 37-42);R973401 (phosphodiesterase Type IV inhibitor; see e.g., Arthritis &Rheumatism (1996) Vol. 39, No. 9 (supplement), S282); MK-966 (COX-2Inhibitor; see e.g., Arthritis & Rheumatism (1996) Vol. 39, No. 9(supplement), S81); Iloprost (see e.g., Arthritis & Rheumatism (1996)Vol. 39, No. 9 (supplement), S82); methotrexate; thalidomide (see e.g.,Arthritis & Rheumatism (1996) Vol. 39, No. 9 (supplement), S282) andthalidomide-related drugs (e.g., Celgen); leflunomide (anti-inflammatoryand cytokine inhibitor; see e.g., Arthritis & Rheumatism (1996) Vol. 39,No. 9 (supplement), S131; Inflammation Research (1996) Vol. 45, pp.103-107); tranexamic acid (inhibitor of plasminogen activation; seee.g., Arthritis & Rheumatism (1996) Vol. 39, No. 9 (supplement), S284);T-614 (cytokine inhibitor; see e.g., Arthritis & Rheumatism (1996) Vol.39, No. 9 (supplement), S282); prostaglandin E1 (see e.g., Arthritis &Rheumatism (1996) Vol. 39, No. 9 (supplement), S282); Tenidap(non-steroidal anti-inflammatory drug; see e.g., Arthritis & Rheumatism(1996) Vol. 39, No. 9 (supplement), S280); Naproxen (non-steroidalanti-inflammatory drug; see e.g., Neuro Report (1996) Vol. 7, pp.1209-1213); Meloxicam (non-steroidal anti-inflammatory drug); Ibuprofen(non-steroidal anti-inflammatory drug); Piroxicam (non-steroidalanti-inflammatory drug); Diclofenac (non-steroidal anti-inflammatorydrug); Indomethacin (non-steroidal anti-inflammatory drug);Sulfasalazine (see e.g., Arthritis & Rheumatism (1996) Vol. 39, No. 9(supplement), S281); Azathioprine (see e.g., Arthritis & Rheumatism(1996) Vol. 39, No. 9 (supplement), S281); ICE inhibitor (inhibitor ofthe enzyme interleukin-1β converting enzyme); zap-70 and/or lckinhibitor (inhibitor of the tyrosine kinase zap-70 or lck); VEGFinhibitor and/or VEGF-R inhibitor (inhibitors of vascular endothelialcell growth factor or vascular endothelial cell growth factor receptor;inhibitors of angiogenesis); corticosteroid anti-inflammatory drugs(e.g., SB203580); TNF-convertase inhibitors; anti-IL-12 antibodies;anti-IL-18 antibodies; interleukin-11 (see e.g., Arthritis & Rheumatism(1996) Vol. 39, No. 9 (supplement), S296); interleukin-13 (see e.g.,Arthritis & Rheumatism (1996) Vol. 39, No. 9 (supplement), S308);interleukin −17 inhibitors (see e.g., Arthritis & Rheumatism (1996) Vol.39, No. 9 (supplement), S120); gold; penicillamine; chloroquine;chlorambucil; hydroxychloroquine; cyclosporine; cyclophosphamide; totallymphoid irradiation; anti-thymocyte globulin; anti-CD4 antibodies;CD5-toxins; orally-administered peptides and collagen; lobenzaritdisodium; Cytokine Regulating Agents (CRAs) HP228 and HP466 (HoughtenPharmaceuticals, Inc.); ICAM-1 antisense phosphorothioateoligo-deoxynucleotides (ISIS 2302; Isis Pharmaceuticals, Inc.); solublecomplement receptor 1 (TP10; T Cell Sciences, Inc.); prednisone;orgotein; glycosaminoglycan polysulphate; minocycline; anti-IL2Rantibodies; marine and botanical lipids (fish and plant seed fattyacids; see e.g., DeLuca et al. (1995) Rheum. Dis. Clin. North Am.21:759-777); auranofin; phenylbutazone; meclofenamic acid; flufenamicacid; intravenous immune globulin; zileuton; azaribine; mycophenolicacid (RS-61443); tacrolimus (FK-506); sirolimus (rapamycin); amiprilose(therafectin); cladribine (2-chlorodeoxyadenosine); methotrexate;antivirals; and immune modulating agents. Any of the above-mentionedagents can be administered in combination with the TNFα inhibitor,including a TNFα antibody, to treat erosive polyarthritis or to inhibitradiographic progression of joint disease.

In one embodiment, the TNFα inhibitor, e.g., a TNFα antibody, orantigen-binding portion thereof, is administered in combination with oneof the following agents for the treatment of rheumatoid arthritis: smallmolecule inhibitor of KDR (ABT-123), small molecule inhibitor of Tie-2;methotrexate; prednisone; celecoxib; folic acid; hydroxychloroquinesulfate; rofecoxib; etanercept; infliximab; leflunomide; naproxen;valdecoxib; sulfasalazine; methylprednisolone; ibuprofen; meloxicam;methylprednisolone acetate; gold sodium thiomalate; aspirin;azathioprine; triamcinolone acetonide; propxyphene napsylate/apap;folate; nabumetone; diclofenac; piroxicam; etodolac; diclofenac sodium;oxaprozin; oxycodone hcl; hydrocodone bitartrate/apap; diclofenacsodium/misoprostol; fentanyl; anakinra, human recombinant; tramadol hcl;salsalate; sulindac; cyanocobalamin/fa/pyridoxine; acetaminophen;alendronate sodium; prednisolone; morphine sulfate; lidocainehydrochloride; indomethacin; glucosamine sulfate/chondroitin;cyclosporine; amitriptyline hcl; sulfadiazine; oxycodonehcl/acetaminophen; olopatadine hcl; misoprostol; naproxen sodium;omeprazole; mycophenolate mofetil; cyclophosphamide; rituximab; IL-1TRAP; MRA; CTLA4-IG; IL-18 BP; ABT-874; ABT-325 (anti-IL 18); anti-IL15; BIRB-796; SCIO-469; VX-702; AMG-548; VX-740; Roflumilast; IC-485;CDC-801; and mesopram. In another embodiment, the TNFα antibody of theinvention is administered for the treatment of a TNFα related disorderin combination with one of the above mentioned agents for the treatmentof rheumatoid arthritis.

In one embodiment, the TNFα inhibitor, e.g., a TNFα antibody, orantigen-binding portion thereof, is used in combination with a drug usedto treat Crohn's disease or a Crohn's-related disorder. Examples oftherapeutic agents which can be used to treat Crohn's includemesalamine, prednisone, azathioprine, mercaptopurine, infliximab,budesonide, sulfasalazine, methylprednisolone sod succ,diphenoxylate/atrop sulf, loperamide hydrochloride, methotrexate,omeprazole, folate, ciprofloxacin/dextrose-water, hydrocodonebitartrate/apap, tetracycline hydrochloride, fluocinonide,metronidazole, thimerosal/boric acid, cholestyramine/sucrose,ciprofloxacin hydrochloride, hyoscyamine sulfate, meperidinehydrochloride, midazolam hydrochloride, oxycodone hcl/acetaminophen,promethazine hydrochloride, sodium phosphate,sulfamethoxazole/trimethoprim, celecoxib, polycarbophil, propoxyphenenapsylate, hydrocortisone, multivitamins, balsalazide disodium, codeinephosphate/apap, colesevelam hcl, cyanocobalamin, folic acid,levofloxacin, methylprednisolone, natalizumab, and interferon-gamma.

In one embodiment, the TNFα inhibitor, e.g., a TNFα antibody, orantigen-binding portion thereof, is administered in combination with anagent which is commonly used to treat spondyloarthropathies, such as AS.Examples of such agents include nonsteroidal, anti-inflammatory drugs(NSAIDs), COX 2 inhibitors, including Celebrex® Vioxx® and Bextra® aandetoricoxib. Physiotherapy is also commonly used to treatspondyloarthropathies, usually in conjunction with non-steoidalinflammatory drugs.

In one embodiment, the TNFα inhibitor, e.g., a TNFα antibody, orantigen-binding portion thereof, is administered in combination with anadditional therapeutic agent to treat ankylosing spondylitis. Examplesof agents which can be used to reduce or inhibit the symptoms ofankylosing spondylitis include ibuprofen, diclofenac and misoprostol,naproxen, meloxicam, indomethacin, diclofenac, celecoxib, rofecoxib,sulfasalazine, prednisone, methotrexate, azathioprine, minocyclin,prednisone, etanercept, and infliximab.

In one embodiment, the TNFα inhibitor, e.g., a TNFα antibody, orantigen-binding portion thereof, is administered in combination with anadditional therapeutic agent to treat psoriatic arthritis. Examples ofagents which can be used to reduce or inhibit the symptoms of psoriaticarthritis include methotrexate; etanercept; rofecoxib; celecoxib; folicacid; sulfasalazine; naproxen; leflunomide; methylprednisolone acetate;indomethacin; hydroxychloroquine sulfate; sulindac; prednisone;betamethasone diprop augmented; infliximab; methotrexate; folate;triamcinolone acetonide; diclofenac; dimethylsulfoxide; piroxicam;diclofenac sodium; ketoprofen; meloxicam; prednisone;methylprednisolone; nabumetone; tolmetin sodium; calcipotriene;cyclosporine; diclofenac; sodium/misoprostol; fluocinonide; glucosaminesulfate; gold sodium thiomalate; hydrocodone; bitartrate/apap;ibuprofen; risedronate sodium; sulfadiazine; thioguanine; valdecoxib;alefacept; and efalizumab.

In one embodiment, the TNFα inhibitor, e.g., a TNFα antibody, orantigen-binding portion thereof, is administered in combination withtopical corticosteroids, vitamin D analogs, and topical or oralretinoids, or combinations thereof, for the treatment of psoriasis. Inaddition, the TNFα antibody of the invention is administered incombination with one of the following agents for the treatment ofpsoriasis: small molecule inhibitor of KDR (ABT-123), small moleculeinhibitor of Tie-2, calcipotriene, clobetasol propionate, triamcinoloneacetonide, halobetasol propionate, tazarotene, methotrexate,fluocinonide, betamethasone diprop augmented, fluocinolone, acetonide,acitretin, tar shampoo, betamethasone valerate, mometasone furoate,ketoconazole, pramoxine/fluocinolone, hydrocortisone valerate,flurandrenolide, urea, betamethasone, clobetasol propionate/emoll,fluticasone propionate, azithromycin, hydrocortisone, moisturizingformula, folic acid, desonide, coal tar, diflorasone diacetate,etanercept, folate, lactic acid, methoxsalen, hc/bismuthsubgal/znox/resor, methylprednisolone acetate, prednisone, sunscreen,salicylic acid, halcinonide, anthralin, clocortolone pivalate, coalextract, coal tar/salicylic acid, coal tar/salicylic acid/sulfur,desoximetasone, diazepam, emollient, pimecrolimus emollient,fluocinonide/emollient, mineral oil/castor oil/na lact, mineraloil/peanut oil, petroleum/isopropyl myristate, psoralen, salicylic acid,soap/tribromsalan, thimerosal/boric acid, celecoxib, infliximab,alefacept, efalizumab, tacrolimus, pimecrolimus, PUVA, UVB, andsulfasalazine.

An antibody, antibody portion, or other TNFα inhibitor, e.g., a TNFαantibody, or antigen-binding portion thereof, may be used in combinationwith other agents to treat skin conditions. For example, an antibody,antibody portion, or other TNFα inhibitor of the invention is combinedwith PUVA therapy. PUVA is a combination of psoralen (P) and long-waveultraviolet radiation (UVA) that is used to treat many different skinconditions. The antibodies, antibody portions, or other TNFα inhibitorsof the invention can also be combined with pimecrolimus. In anotherembodiment, the antibodies of the invention are used to treat psoriasis,wherein the antibodies are administered in combination with tacrolimus.In a further embodiment, tacrolimus and TNFα inhibitors are administeredin combination with methotrexate and/or cyclosporine. In still anotherembodiment, the TNFα inhibitor of the invention is administered withexcimer laser treatment for treating psoriasis.

Nonlimiting examples of other therapeutic agents with which a TNFαinhibitor, e.g., a TNFα antibody, or antigen-binding portion thereof,can be combined to treat a skin or nail disorder include UVA and UVBphototherapy. Other nonlimiting examples which can be used incombination with a TNFα inhibitor include anti-IL-12 and anti-IL-18therapeutic agents, including antibodies.

Any one of the above-mentioned therapeutic agents, alone or incombination therewith, can be administered to a subject suffering fromerosive polyarthritis, in combination with the TNFα a inhibitor,including a TNFα antibody, or antigen-binding portion thereof. A TNFαantibody, or antigen-binding portion thereof, may be used in combinationwith additional therapeutic agents known to be effective at acutemanagement of subjects with erosive polyarthritis. A TNFα antibody, orantigen-binding portion thereof, may also be used in combination withadditional therapeutic agents known to be effective at management ofsubjects with erosive polyarthritis.

This invention is further illustrated by the following example whichshould not be construed as limiting. The contents of all references,patents and published patent applications cited throughout thisapplication are incorporated herein by reference.

EXAMPLE Treatment of Erosive Polyarthritis in Patients with PsoriaticArthritis Using a TNF Inhibitor

Erosive polyarthritis occurs in a substantial proportion of patientswith psoriatic arthritis (PsA). Traditional, non-biologic DMARDs havenot been shown to effectively inhibit the radiographic progression ofjoint damage in this disease.

The following study was performed to evaluate the efficacy of a TNFinhibitor, more specifically the anti-TNF antibody adalimumab, for thetreatment of erosive polyarthritis. The study was performed to evaluatewhether adalimumab was effective at inhibiting the radiographicprogression of joint disease associated with erosive polyarthritis inpatients with moderate to severe PsA.

A 24-week, double-blind, randomized, placebo-controlled trial of adultpatients with moderate to severely active PsA (≧3 swollen and ≧3 tenderjoints) who had failed NSAID therapy was performed. Patients werestratified according to methotrexate (MTX) use (yes/no) and degree ofpsoriasis (<3% or ≧3% Body Surface Area [BSA]). In addition to having ≧3swollen and ≧3 tender joints, inclusion criteria included an inadequateresponse to NSAID therapy, a history of psoriasis, and age ≧18 years.Exclusion criteria included the following: prior anti-TNF therapy,alefacept within 12 weeks prior to study entry, other biologics within 6weeks prior to study entry, systemic therapies for psoriasis within 4weeks prior to study entry, and phototherapy and topicals within 2 weeksprior to study entry.

Patients were randomized to receive either adalimumab 40 mg or placebosubcutaneously every other week (eow) for 24 weeks. Patients whocompleted the 24-week trial were eligible to enroll in an open-labelextension (OLE) study, in which all patients received adalimumab eow.After 12 weeks of treatment with open label therapy, patients failing tomeet pre-specified criteria were eligible to receive 40 mg weekly.

Radiographic assessments were performed during both the blinded portion(Weeks 0 and 24) and the open-label portion (Week 48). Radiographs ofthe hands and feet were assessed by a modified total Sharp score (mTSS)in which additional joints typically involved in PsA were added, and, tobetter quantify the significant osteolysis that occurs in PsA, thenumerical scales were expanded. The mTSS was determined by combining thejoint space narrowing score (0-192) and the erosion score (0-378), asshown in FIG. 1 a. Clinical findings associated with PsA, e.g.,pencil-in-cup changes, were also evaluated. Diagrams of the mTSS and theradiographic findings associated with PsA used in this study are shownin FIGS. 1 a and 1 b, respectively.

The radiograph reading procedure included the following. All films wereread by two independent readers who were blinded to treatment and filmorder. Read number 1 was an evaluation of baseline and week 24 films.Read number 2 was an evaluation of baseline, week 24, and week 48 films.

Several sensitivity analyses were utilized to assess the impact ofmissing radiographs (imputation of zero change, worst rank change,50^(th)/75^(th) percentile change based on patients with similarbaseline scores, and linear extrapolation when multiple radiographs wereavailable).

Week 24 analysis included the following: inclusion in the week 24analysis required both baseline and week 24 films, where at least 50% ofthe joints were evaluable. Week 48 analysis included the following: Allpatients from the 24 week analysis were included in the week 48analysis. If the week 48 film was not available (or <50% of the jointsevaluable), then the following imputation was performed: if originallyrandomized to placebo, a change of 0 was imputed; and if originallyrandomized to adalimumab, linear extrapolation using first two films wasconducted.

Baseline demographics and disease severity characteristics wereconsistent with moderate to severe PsA and were well-matched betweentreatment arms (adalimumab N=151, placebo N=162; mean±SD): age 49.0±11.8yrs; duration of PsA 9.5±8.5 yrs; SJC (76) 14±12; TJC (78) 25±18; HAQ1.0±0.6; mTSS 20.8±40.9; 51% were taking concomitant methotrexate. Ofthe total, 296 patients had X-rays at baseline and Week 24, and 265patients also had X-rays at Week 48.

As reported in previous studies, the ACR20, 50, and 70 responses and thePASI 50, 75, and 90 responses for adalimumab-treated patients at week 24were significantly better than placebo. ACR and PASI responses at week24 are shown below in tables 1 and 2 (all results p≦0.001 placebo vs.adalimumab):

TABLE 1 ACR response: % of patients ACR20 ACR50 ACR70 Placebo (N = 162)15 6 1 Adalimumab (N = 151) 57 39 23

TABLE 2 PASI response: % of patients PASI 50 PASI 75 PASI 90 Placebo (N= 69) 12 1 0 Adalimumab (N = 69) 75 59 42

During the blinded study period (24 wks), adalimumab patients hadsignificantly less progression in mTSS than placebo patients (meanchange in mTSS −0.2 vs. 1.0, p<0.001, ranked ANCOVA). Statisticalsignificance was maintained in all sensitivity analyses. FIG. 2 showsthe distribution of mTSS scores which demonstrates that fewer patientstreated with adalimumab had an increase in structural damage during 24weeks of treatment compared with placebo. The difference in distributionwas observed by looking at mean scores at week 24 and the number andpercentage of patients who had an increase in Sharp score during thestudy (see Table 3). Approximately three times as many placebo-treatedpatients had an increase in mTSS (>0.5 units) than adalimumab-treatedpatients during the first 24 weeks of treatment.

TABLE 3 Change* in Modified Total Sharp Score at Week 24 PlaceboAdalimumab N = 152 N = 144 n (%) n (%) Decrease in Sharp Score 8 (5.3%)27 (18.8%) No change in Sharp Score 100 (65.8%) 104 (72.2%) Increase inSharp Score 44 (28.9%) 13 (9.0%) p ≦ 0.001 placebo vs. adalimumab usingCMH test *Change defined as >0.5 unites in mTSS Score

Statistically significant differences were observed between adalimumaband placebo treated subjects for both erosion scores and joint narrowingscores (p≦0.001 using a ranked ANACOVA). At Week 24, the change inerosion scores (change from baseline) were 0.6 for placebo patients and0.0 for adalimumab patients (p<0.001, ranked ANCOVA), and the change injoint space narrowing scores (change from baseline) were 0.4 for placebopatients and −0.2 for adalimumab patients (p<0.001, ranked ANCOVA).

Sensitivity analyses to account for missing patients films wereperformed and results maintained statistical significance with allanalyses. Post-hoc sensitivity analyses excluding feet and DIPs were asfollows: one analysis was run excluding feet and DIPs and a secondanalysis was run excluding all DIP joints. Statistical significance wasmaintained in both analyses.

Statistically significant differences were observed between adalimumaband placebo treated subjects regardless of whether concomitant MTX wasbeing used. Mean differences were slightly higher for patients takingconcomitant MTX. Patients on monotherapy showed a change from baselineof −0.1 in adalimumab (n=68) vs. 0.9 for placebo (n=74) (p≦0.001 using aranked ANACOVA). Patients on concomitant MTX showed a change frombaseline of −0.3 in adalimumab (n=76) vs. 1.2 for placebo (n=78)(p≦0.001 using a ranked ANACOVA). FIGS. 3 a and 3 b show cumulativedistribution function plots of mTSS of subjects with and without MTX.

Analysis of 48-week radiographs demonstrated that the lack ofprogression (lack of changes in mTSS) observed at Week 24 was maintainedto Week 48 in adalimumab patients (see FIG. 4). Patients treated withplacebo for 24 weeks did not have radiographic progression of diseaseduring the open-label period. Neither treatment arm demonstratedsignificant progression in PsA-associated features. The prevalence ofPsA-associated findings are shown in Table 4. No significant differencewas found between groups at baseline, and no significant progression wasfound in either group during the 24-week study.

TABLE 4 Prevalence of PsA-associated findings All patients (N = 313) n(%) Joint space widening 38 (12.1%) Gross osteolysis 60 (19.2%)Subluxation 49 (15.7%) Pencil-in-cup 9 (2.9%) Juxta-articularperiostitis 247 (78.9%) Shaft periostitis 140 (44.7%) Phalangeal tuftresorption 224 (71.6%)

Furthermore, adalimumab was generally well-tolerated as reportedpreviously.

Adalimumab was more effective compared with placebo in inhibitingradiographic disease progression over a 24-week period. Adalimumabshowed differences versus placebo both in patients taking concomitantmethotrexate and in those taking adalimumab as monotherapy. Theinhibition of structural damage progression observed inadalimumab-treated patients at 24 weeks was maintained at one year. Inconclusion, this study demonstrated that adalimumab was effective intreating erosive polyarthritis and radiographic disease progression overone year in patients who also had moderate to severely active PsA.

Multiple-Variable Dose Treatment of Psoriasis With D2E7

A study was performed to determine the efficacy of a multiple-variabledose regimen of D2E7 for treating psoriasis. Efficacy and tolerabilityof D2E7 in the treatment of patients with moderate to severe chronicplaque psoriasis were evaluated in a randomized, double-blind,placebo-controlled multicenter study.

In this study, one hundred forty-eight adult patients with a diagnosisof moderate to severe psoriasis for at least one year were selected toreceive multiple-variable dose treatment. Patients were also selectedbased on an affected body surface area (BSA) of ≧5%. Subjects wererandomized equally to one of three groups (two treatment groups and oneplacebo).

At baseline (Week 0) patients in both treatment groups received aninduction dose of 80 mg of D2E7. Patients in the first treatment groupsubsequently received a treatment dose of 40 mg of D2E7 at week 1followed by 40 mg every other week (eow) starting at week 3. Subjects inthe second treatment group received an induction dose of 80 mg dose ofD2E7 at Week 1 (following the initial 80 mg dose at Week 0) followed bya treatment dose of 40 mg of D2E7 weekly starting at week 2. The placebogroup received only the placebo weekly starting at baseline. Alltreatment was administered subcutaneously (sc) with pre-filled syringes.A summary of the different regimens are described below in Table 2:

TABLE 2 Psoriasis study regimens Regimen Detailed description A D2E7 80mg sc administered at Week 0 (baseline); D2E7 40 mg sc every other weekadministered starting at Week 1 through Week 11, with placeboadministered on alternate weeks B D2E7 80 mg sc administered starting atWeek 0 (baseline) and at Week 1(80/80); D2E7 40 mg sc weeklyadministered starting at Week 2 through Week 11 C Placebo sc will beadministered at baseline and then weekly through Week 11, with twoinjections given at Week 1 and Week 1

In order to maintain the blind study, all subjects received a total of 2injections at baseline and week 1. During the remaining period of thestudy (weeks 2 through 12), subjects received one injection per week.The treatment dose per injection correlated to the dose regimen randomlyassigned to each subject.

The PASI of the participants of the multiple-variable dose regimen wasdetermined according to standard methods (see Fredriksson andPettersson, supra and Marls et al., supra). The primary efficacyendpoint of the study was the percentage of subjects achieving aclinical response as defined by at least a 75% reduction in the PASIscore (≧PAST 75) at Week 12.

Secondary efficacy measures included a static Physician's GlobalAssessment (PGA) of “clear” or “almost clear” at Week 12. PGA wasdetermined according to a seven point scale used to measure the severityof psoriasis at the time of the physician's evaluation. Descriptions ofthe disease used included the following: severe=very marked plaqueelevation, scaling, and/or erythema; moderate to severe=marked plaqueelevation, scaling, and/or erythema; moderate=moderate plaque elevation,scaling, and/or erythema; mild to moderate=intermediate between moderateand mild; mild=slight plaque elevation, scaling, and/or erythema; almostclear=intermediate between mild and clear; and clear=no signs ofpsoriasis.

The results show that at Week 12, statistically significantly greaterpercentages of patients achieved a PASI 75 response or better on D2E7than those on a placebo treatment. For patients receiving 40 mgtreatment dose of D2E7 eow, 53% demonstrated a PASI of 75 or higher. Inaddition, 80% of patients receiving a 40 mg treatment dose of D2E7weekly showed a PASI 75 or higher, compared to only 4% of the placebotreatment group (p<0.001 vs. placebo). Response rates at Week 12 forboth dosing regimens of D2E7 were statistically significantly greaterthan for placebo, as shown in FIG. 5.

Overall, the mean percentage changes in PASI score for patients on D2E7multiple-variable dose therapy were statistically significantly greaterthan placebo. The changes were evident as early as Week 1 after theinitial dose, as shown in FIG. 6. At Week 12, 49% of patients on D2E7receiving 40 mg eow and 76% of patients receiving 40 mg of D2E7 weeklyachieved a PGA of “clear” or “almost clear,” compared with 2% of placebopatients.

Overall, the mean percentage changes in PASI score for patients on D2E7multiple-variable dose therapy were statistically significantly greaterthan placebo. The changes were evident as early as Week 1 after theinitial dose, as shown in FIG. 6. At Week 12, 49% of patients on D2E7receiving 40 mg eow and 76% of patients receiving 40 mg of D2E7 weeklyachieved a PGA of “clear” or “almost clear,” compared with 2% of placebopatients.

Of the one hundred forty-eight adult patients enrolled in the study, 29%also had a medical history of psoriatic arthritis (PsA). Both doses ofD2E7 were effective in the treatment of psoriasis in both patients withand without PsA. Patients with PsA had a similar efficacy response toD2E7 as those without PsA. For both PsA and without PsA subgroups, thepercentages of patients achieving a PASI 75 response or better at week12 was statistically significant for the eow (with PsA, 47%; withoutPsA, 57%) and weekly treatment arms (with PsA, 58%; without PsA, 87%)compared with placebo. Continued improvements in efficacy were seenthrough week 24 in the eow arm (with PsA, 53%; without PsA, 70%).Efficacy responses in patients with and without PsA at weeks 12 and 24are shown in FIGS. 7 and 8, respectively.

In conclusion, D2E7 administered for 12 weeks was effective in thetreatment of moderate to severe chronic plaque psoriasis. 53% ofpatients on 40 mg eow achieved ≧PASI 75, compared with 4% on placebo.80% of patients on 40 mg weekly achieved ≧PASI 75.49% and 76% ofpatients on D2E7 40 mg eow and 40 mg weekly, respectively, were “clear”or “almost clear” of their psoriasis. In addition, D2E7 was equallyeffective at treating psoriasis patients with and without PsA.

EQUIVALENTS

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, many equivalents to the specificembodiments of the invention described herein. Such equivalents areintended to be encompassed by the following claims. The contents of allreferences, patents and published patent applications cited throughoutthis application are incorporated herein by reference

What is claimed is:
 1. A method for treating moderate to severe chronicplaque psoriasis, comprising subcutaneously administering to an adultpatient having moderate to severe chronic plaque psoriasis a first doseof 80 mg of adalimumab, followed by 40 mg of adalimumab every other weekstarting one week after said first dosing, wherein the patient achievesat least Psoriasis Area and Severity Index (PASI) 90 response at week 12of the treatment.
 2. The method of claim 1, wherein adalimumab iscomprised in pre-filled syringes for subcutaneous injection.
 3. A methodfor treating moderate to severe chronic plaque psoriasis, comprisingsubcutaneously administering to an adult patient having moderate tosevere chronic plaque psoriasis an initial dose of 80 mg of adalimumab,followed by 40 mg of adalimumab every other week starting one week aftersaid initial dosing, wherein the patient achieves at least PASI 75response at week 24 of the treatment.
 4. The method of claim 3, whereinadalimumab is comprised in pre-filled syringes for subcutaneousinjection.
 5. A method for treating moderate to severe chronic plaquepsoriasis, comprising subcutaneously administering to an adult patienthaving moderate to severe chronic plaque psoriasis an initial dose of 80mg of adalimumab, followed by 40 mg of adalimumab every other weekstarting one week after said initial dosing, wherein when said method isused to treat a population of patients with moderate to severe chronicplaque psoriasis, at least 47% of said patients achieve at least PASI 75response at week 12 of the treatment.
 6. The method of claim 5, whereinadalimumab is comprised in pre-filled syringes for subcutaneousinjection.
 7. A method for treating moderate to severe chronic plaquepsoriasis, comprising subcutaneously administering to an adult patienthaving moderate to severe chronic plaque psoriasis an initial dose of 80mg of adalimumab, followed by 40 mg of adalimumab every other weekstarting one week after said initial dosing, wherein when said method isused to treat a population of patients with moderate to severe chronicplaque psoriasis, at least 53% of said patients achieve at least PASI 75response at week 24 of the treatment.
 8. The method of claim 7, whereinadalimumab is comprised in pre-filled syringes for subcutaneousinjection.